FORM 2
THE PATENTS ACT, 1970 (39 of
1970)
&
The Patent Rules, 2003
COMPLETE SPECIFICATION
(See section 10 and rule 13)
STABLE CYCLOSPORINE OPHTHALMIC FORMULATION AND
MANUFACTURING PROCESS THEREOF
SUN PHARMACEUTICAL INDUSTRIES LIMITED
A company incorporated under the Laws of India having their corporate office at
SUN HOUSE,201 B/1, Western Express Highway, Goregaon (E), Mumbai -400063,
Maharashtra, India.
The following specification particularly describes the nature of this invention and the
mannerin which it is to be performed:
2
CROSS REFERENCE
The present patent application is a national phase of PCT/IB2022/057851which claims the
benefit of the priority date of Indian Provisional Patent Application No. 202121037917
filed on August 20, 2021.
FIELD OF THE INVENTION
The present invention relates to a stable nanomicellar ophthalmic solution
5 comprising cyclosporine and a method of preparing the nanomicellar solution. The present
invention further relates to the stable nanomicellar solution comprising a cyclosporine form
with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9 or amorphous
cyclosporine. The present invention further relates to the stable nanomicellar solution
comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7,
10 14.4 and 17.5 or a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 8.5,
9.3, 11.6 and 20.3. The present invention also relates to use of this stable nanomicellar
ophthalmic solution in dry eye.
BACKGROUND OF THE INVENTION
Cyclosporine nanomicellar ophthalmic solutions are generally disclosed in U.S.
15 Patent No. 10,918,694, wherein the ophthalmic solution comprises comprising 0.087-0.093
wt% cyclosporine, a polyoxyl lipid or fatty acid and a polyalkoxylated alcohol. Preferably,
it comprises 0.087-0.093 wt% cyclosporine, 0.5-5% of one or more selected from the
group consisting ofHCO-40, HCO-60, HCO-80 and HCO-100; and about 0.01-0.1%
octoxynol-40. Further, it discloses methods of preparing such cyclosporine solutions.
20 The method includes dissolution of cyclosporine in polyoxyl castor oil such as
hydrogenated castor oil and a polyalkoxylated alcohol such as octoxynol at 60°C prior to
addition in an aqueous phase. Specifically, the preparation method of ophthalmic solution
consists of the following steps: HCO-40 is melted in a flask heated to about 60°C with
stirring. When liquefied, the required amount of cyclosporine is added and mixed until
25 dissolved and uniform. Then, octoxynol-40 is heated to about 60°C and when liquefied, is
added to the cyclosporine HCO-40 mixture. Water for injection at about 25°C is
charged into the flask containing the dissolved cyclosporine and stirred until dissolved.
Other excipients are then added, such as sodium chloride and phosphate buffer and then
PVP-K90 and mixed until dissolved and brought to the final volume with water for
30 injection. However, current methods have a problem in that when cyclosporine is
dissolved in the polyoxyl castor oil, such asHCO-40, due to the difference in solubility and
stability of different forms of cyclosporine, there may
3
be a difference in stability ofdifferent batches during manufacturing. The result is that some
batches are not stable and the cyclosporine precipitates out of solution.
Further, cyclosporine in different forms shows different solubility and stability.
Cyclosporine with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9 is the
5 most soluble form of cyclosporine and is useful in the preparation of solution formulations,
however this is not the most stable form and it may convert to the less soluble forms of
cyclosporine, thus affecting the stability of the solution. This form may convert to a more
stable and less soluble cyclosporine with characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7,
14.4 and 17.5 during dissolution of cyclosporine in surfactants at 55-60°C. Similarly, it may
10 change to another less soluble form of cyclosporine with characteristic XRD peaks at 2-
theta (deg.) 8.5, 9.3, 11.6 and 20.3. Further, the amorphous form, which is also one of the
more soluble forms and useful in the preparation of solution formulations, may also
recrystallize to these two less soluble forms. Such a conversion is facilitated by several
factors not limited to moisture, water, solvent temperature and so on. This conversion also
15 depends on stresses like temperature, long storage at higher temperature, and the like. The
process needs modification depending on the form of cyclosporine used, such that the
amorphous form and the form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9 needs wetting followed by complete dissolution at the temperature up to 70°C.
However, if the less soluble forms are used, temperatures as high as 130°C are needed.
20 Further, during the manufacturing steps, due to the instability and conversions, the solution
is to be manufactured with tight control of API specification, process temperature and time
to avoid interconversion of forms. It is rather, difficult to identify to what extent this
conversion has taken place before completing the manufacturing process to get the
nanomicellar cyclosporine. This leads to a formation ofseeds within the composition either
25 at the initial stage of the process or during storage at higher temperatures, which later
crystallized out from the drug product rendering the product unuseful. The soft mesophasic
or liquid crystalline form of cyclosporine formed as intermediate in the non-aqueous phase
may be responsible for such instability.
Thus, there is a need for a stable formulation and method ofits preparation to prevent
30 conversion to less soluble forms of cyclosporine during the manufacturing process and
during storage. The present invention discloses a stable nanomicellar ophthalmic
formulation and an improved method of making such stable formulation. The method for
making the formulation results in a stable formulation irrespective of any form of
4
cyclosporine being used in the formulation. The method does not lead to conversion of one
form to another. More specifically, the present method does not lead to conversion of the
soluble form of cyclosporine to the less soluble forms of cyclosporine and further prevents
precipitation of cyclosporine in the formulation during long-term stability.
5 SUMMARY OF THE INVENTION
One of the objectives of the present invention, according to some embodiments, is a
method of making a stable nanomicellar ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine, adding octoxynol-40; and
c) mixing the resulting mixture with the aqueous vehicle.
The present inventors have surprisingly found that the solution stability of
cyclosporine A at 35°C-40°C was higher than that of 55°C-60°C and thus, reducing the
temperature to 35°C-40°C overcomes the above mentioned stability concerns of the
formulation and provides for a more stable formulation.
In another aspect, the present invention is drawn to a stable nanomicellar ophthalmic
20 formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
25 b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine, adding octoxynol-40; and
c) mixing the resulting mixture with the aqueous vehicle.
In one aspect, mixing the aqueous vehicle occurs at a temperature of 35°C±2°C. In
another aspect, mixing the aqueous vehicle occurs at a temperature of 55±2°C.
5
In one aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
6.9, 7.8, 9.4 and 15.9.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
5 formulation comprising an amorphous form of cyclosporine.
In another aspect, the present invention discloses a method of making a stable
nanomicellar ophthalmic formulation comprising: cyclosporine, hydrogenated 40 polyoxyl
castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) keeping mixture A under vacuum to remove foam;
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine;
15 d) adding octoxynol-40; and
e) mixing the resulting mixture with the aqueous vehicle.
In another embodiment, the present invention provides a stable nanomicellar
ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
20 wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) keeping mixture A under vacuum to remove foam;
c) optionally, lowering the temperature of mixture A to a temperature of
25 35°C±2°C prior to the complete dissolution of the cyclosporine;
d) adding octoxynol-40; and
e) mixing the resulting mixture with the aqueous vehicle.
It was surprisingly found that while keeping the mixture under vacuum, the bubbles
are dragged up and accumulate on the surface from where bubbles were gradually removed
6
WO 2023/021492 PCT/IB2022/057851
5
and the bottom portion is cleared up. So dissolution under vacuum or removal of foams
intermittently during dissolution leads to faster dissolution and may not require the lowering
of the temperature of the mixture. Thus, overcoming the above mentioned stability concerns
of the formulation and provides for a more stable formulation.
5 In one aspect, the present invention discloses mixing the aqueous vehicle at a
temperature of 35°C±2°C. In another aspect, the present invention discloses mixing the
aqueous vehicle at a temperature of 55±2°C.
In one aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
10 6.9, 7.8, 9.4 and 15.9.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising an amorphous form of cyclosporine.
In yet another aspect, the present invention provides a stable nanomicellar
ophthalmic formulation comprising:
15 cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
20 c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
In one aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
6.9, 7.8, 9.4 and 15.9.
25 In another aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising an amorphous form of cyclosporine.
In yet another aspect, the present invention provides a stable nanomicellar
ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
7
WO 2023/021492 PCT/IB2022/057851
6
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
5 c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C,
wherein cyclosporine is present in a form having characteristic XRD peaks at 2-theta (deg.)
7.4, 8.7, 14.4 and 17.5.
In yet another aspect, the present invention provides a stable nanomicellar
10 ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
15 b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of
127-130°C,
wherein cyclosporine is present in a form with characteristic XRD peaks at 2-theta (deg.)
8.5, 9.3, 11.6 and 20.3.
20 In yet another aspect, the present invention provides a stable nanomicellar
ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a
25 temperature of 127-130°C to form a mixture A;
b) adding cyclosporine to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
8
In one aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
6.9, 7.8, 9.4 and 15.9.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
5 formulation comprising an amorphous form of cyclosporine.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine in a form having characteristic XRD peaks at 2-theta
(deg.) 7.4, 8.7, 14.4 and 17.5.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
10 formulation comprising a cyclosporine in a form having characteristic XRD peaks at 2-theta
(deg.) 8.5, 9.3, 11.6 and 20.3.
In a preferred aspect, the present invention provides a method of making a stable
nanomicellar ophthalmic formulation comprising:
0.09 wt% cyclosporine,
15 about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
wherein the method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
20 temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine, adding octoxynol-40; and
c) mixing the resulting mixture with the aqueous vehicle.
In one aspect, the aqueous vehicle is mixed at a temperature of 35°C±2°C. In
25 another aspect, the aqueous vehicle is mixed at a temperature of 55±2°C.
In a preferred aspect, the present invention also provides a stable nanomicellar
ophthalmic formulation prepared by a method as described above.
In a preferred aspect, the present invention provides a method of making a stable
nanomicellar ophthalmic formulation comprising:
9
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
5 wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) keeping mixture A under vacuum to remove foam;
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine;
d) adding octoxynol-40; and
e) mixing the resulting mixture with the aqueous vehicle.
In one aspect, the aqueous vehicle is mixed at a temperature of 35°C±2°C. In
another aspect, the aqueous vehicle is mixed at a temperature of 55±2°C.
15 In a preferred aspect, the present invention also provides a stable nanomicellar
ophthalmic formulation prepared by a method as described above.
In one aspect, the stable nanomicellar ophthalmic formulation further comprises:
about 0.20-0.550 wt% sodium phosphate monobasic,
about 0.23-0.465 wt% sodium phosphate dibasic,
20 about 0.05 wt% sodium chloride,
about 0.3 wt% povidone,
sodium hydroxide/hydrochloric acid to adjust the pH, and
water for injection.
In one aspect, the present invention provides a stable nanomicellar ophthalmic
25 formulation, wherein the pH of the formulation is about 5.0 to 8.0. More preferably, the pH
of the formulation is about 6.5 to 7.2.
10
Further, the present invention provides a stable nanomicellar ophthalmic
formulation wherein the osmolality of the formulation is between about 150 to about 200
mOsmol/kg.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
5 formulation comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
6.9, 7.8, 9.4 and 15.9.
In yet another aspect, the present invention provides a stable nanomicellar
ophthalmic formulation comprising an amorphous form of cyclosporine.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
10 formulation, wherein the formulation is substantially free of a cyclosporine form with
characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7, 14.4 and 17.5.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
formulation, wherein the formulation is substantially free of a cyclosporine form with
characteristic XRD peaks at 2-theta (deg.) 8.5, 9.3, 11.6 and 20.3.
15 In another aspect, the present invention provides a stable nanomicellar ophthalmic
formulation, prepared by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
20 to the complete dissolution of the cyclosporine, adding octoxynol-40; and
c) then mixing the resulting mixture with an aqueous vehicle at 35°C±2°C,
wherein the mixture A is lowered to a temperature of 35°C±2°C in less than 65
minutes. Preferably, the mixture A is lowered to a temperature of 35°C±2°C in 40-
50 minutes.
25 In one aspect, the present invention discloses mixing the mixture A of step (a) for
20-30 minutes, preferably, 20-25 minutes, more preferably, for 20 ±2 minutes.
In one aspect, the present invention discloses lowering the temperature to 35°C±2°C
and stirring for 60-70 minutes. Preferably, stirring the mixture for 60±5 minutes at a
temperature of 35°C±2°C.
11
In yet another aspect, the present invention provides a stable nanomicellar
ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
5 about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
In one aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
15 6.9, 7.8, 9.4 and 15.9.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising an amorphous form of cyclosporine.
In yet another aspect, the present invention provides a stable nanomicellar
ophthalmic formulation comprising:
20 0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
25 a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
12
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C,
wherein cyclosporine is present in a form having characteristic XRD peaks at 2-theta (deg.)
7.4, 8.7, 14.4 and 17.5.
5 In yet another aspect, drawn to a stable nanomicellar ophthalmic formulation
compnsmg:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
15 c) mixing the resulting mixture with the aqueous vehicle at a temperature of
127-130°C,
wherein cyclosporine is present in a form with characteristic XRD peaks at 2-theta (deg.)
8.5, 9.3, 11.6 and 20.3.
In yet another aspect, the present invention provides a stable nanomicellar
20 ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
25 wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a
temperature of 127-130°C to form a mixture A;
b) adding cyclosporine to mixture A at 127-130°C; and
13
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
In one aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
5 6.9, 7.8, 9.4 and 15.9.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising an amorphous form of cyclosporine.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine in a form having characteristic XRD peaks at 2-theta
10 (deg.) 7.4, 8.7, 14.4 and 17.5.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine in a form having characteristic XRD peaks at 2-theta
(deg.) 8.5, 9.3, 11.6 and 20.3.
In yet another aspect, the present invention provides a method of making a stable
15 nanomicellar ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40,
about 0.20-0.550 wt% sodium phosphate monobasic,
20 about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
about 0.3 wt% povidone,
sodium hydroxide/hydrochloric acid to adjust the pH, and
water for injection,
25 wherein the method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
14
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine, adding octoxynol-40 to form an API
mixture;
c) adding API mixture to Water for Injection (WFI);
5 d) adding remaining excipients to step (c) in the order of sodium phosphate
monobasic, then sodium phosphate dibasic, then sodium chloride, and then
polyvinylpyrrolidone;
e) adjusting the pH to 6.5 to 7.2 and bringing to the final volume with WFI.
In one aspect, the present invention discloses mixing the mixture A of step (a) for
10 20-30 minutes, preferably, 20-25 minutes, more preferably, for 20 ±2 minutes.
In one aspect, the present invention discloses lowering the temperature of the
mixture A to 35°C±2°C in less than 65 minutes. Preferably, lowering the temperature of the
mixture A to 35°C±2°C in 40-50 minutes.
In another aspect, the present invention discloses stirring the mixture of step (b) for
15 60-70 minutes. Preferably, stirring the mixture for 60±5 minutes at a temperature of
35°C±2°C.
In one aspect, the present invention discloses mixing the API mixture to WFI at a
temperature of35°C±2°C. In another aspect, the present invention discloses mixing the API
mixture to WFI at a temperature of 55±2°C.
20 In yet another aspect, the present invention provides a method of making a stable
nanomicellar ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40,
25 about 0.20-0.550 wt% sodium phosphate monobasic,
about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
about 0.3 wt% povidone,
sodium hydroxide/hydrochloric acid to adjust the pH, and
15
water for injection,
wherein the method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
5 b) keeping mixture A under vacuum to remove foam;
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine;
d) adding octoxynol-40 to form an API mixture;
e) adding the API mixture to Water for Injection (WFI);
f) adding remaining excipients to step (c) in the order of sodium phosphate
monobasic, then sodium phosphate dibasic, then sodium chloride, and then
polyvinylpyrrolidone; and
g) adjusting the pH to 6.5 to 7.2 and bringing to the final volume with WFI.
In one aspect, the present invention discloses mixing the mixture A of step (a) for
15 20-30 minutes, preferably, 20-25 minutes, more preferably, 20 ±2 minutes.
In one aspect, the present invention discloses lowering the temperature of the
mixture A to 35°C±2°C in less than 65 minutes. Preferably, lowering the temperature of the
mixture A to 35°C±2°C in 40-50 minutes.
In one aspect, the present invention discloses mixing the API mixture to WFI at a
20 temperature of 35°C±2°C. In another aspect, mixing the API mixture to WFI occurs at a
temperature of 55±2°C.
In a more preferred aspect, the present invention provides for a stable nanomicellar
ophthalmic formulation prepared by any of the method as described above.
BRIEF DESCRIPTION OF THE DRAWINGS
25 Figure l(a) and Figure l(b) depict cyclosporine nanomicellar ophthalmic
formulations prepared by methods described in Example l(a) and Example l(b),
respectively. Figure l(a) discloses that no particles were observed, while Figure l(b)
discloses that particles were observed on stability.
16
Figure 2 depicts characteristic X-ray powder diffraction (XRPD) patterns of
cyclosporine with a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9,
7.8, 9.4 and 15.9 (A), a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
7.4, 8.7, 14.4 and 17.5 (B), a cyclosporine form with characteristic XRD peaks at 2-theta
5 (deg.) 8.5, 9.3, 11.6 and 20.3 (C), and an amorphous form of cyclosporine.
Figures 3(a) to 3(d) depict the solubility behavior of different cyclosporine forms at
55°C. Figure 3(a) depicts the solubility behavior of CsA form with characteristic XRD peaks
at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9. Figure 3(b) depicts solubility behavior ofa CsA form
with characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7, 14.4 and 17.5. Figure 3(c) depicts
10 solubility behaviour of a CsA form with characteristic XRD peaks at 2-theta (deg.) 8.5, 9.3,
11.6 and 20.3, and Figure 3(d) depicts solubility behaviour of amorphous CsA.
Figure 4 depicts X-ray powder diffraction (XRPD) patterns of a precipitated
cyclosporine from solution prepared when cyclosporine is dissolved in Kolliphor® RH 40
at higher exposure and temperature.
15 Figure 5 depicts X-ray powder diffraction (XRPD) patterns of a precipitated
cyclosporine when cyclosporine is dissolved in Kolliphor® RH 40 and the kept for a longer
time till the precipitation occurred.
DETAILED DESCRIPTION OF THE INVENTION
One embodiment of the present disclosure is a stable nanomicellar ophthalmic
20 formulation. The ophthalmic formulation comprises cyclosporine, a polyoxyl lipid or fatty
acid, and a polyalkoxylated alcohol.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising:
cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated alcohol, wherein the
25 ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the polyoxyl lipid or fatty acid at a temperature
of 55°C or above to form a mixture A; and
b) lowering the temperature of mixture A to a temperature not higher than 40°C
prior to the complete dissolution of the cyclosporine.
30 Preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is done at a
temperature of 55°C-60°C to form a mixture A. Preferably, mixture A is lowered to a
17
temperature of 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is done at a
temperature of 55°C±2°C to form a mixture A; and mixture A is lowered to a temperature
not higher than 35°C±2°C prior to the complete dissolution of the cyclosporine. Further, the
5 method comprises adding the polyalkoxylated alcohol and then mixing the resulting mixture
with an aqueous vehicle at 35°C±2°C. In another aspect, the resulting mixture with an
aqueous vehicle at a temperature of 55±2°C.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising:
10 cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated alcohol, and an aqueous
vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the polyoxyl lipid or fatty acid at a temperature
of 55°C±2°C or above to form a mixture A;
15 b) keeping mixture A under vacuum to remove foam;
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine, adding
polyalkoxylated alcohol; and
d) mixing the resulting mixture with the aqueous vehicle.
20 In one aspect, the aqueous vehicle is mixed at a temperature of 35°C±2°C. In
another aspect, the aqueous vehicle is mixed at a temperature of 55±2°C.
It was found that during the dissolution of hydrophobic molecule like cyclosporine
A (CsA) in polyoxyl lipid, such as, polyoxyl hydrogenated castor oil, that air was entrapped
in the bulk in a form of bubbles and creates foam. This happens irrespective of temperature
25 during the drug dissolution process, either 55°C or even when the temperature was reduced
to 35°C. This may delay the wetting and dissolution of cyclosporine as this entrapped air
may make a boundary between cyclosporine particles and water. It was surprisingly found
that by keeping the mixture under vacuum, the bubbles are dragged up and accumulate on
the surface from where bubbles were gradually removed and the bottom portion is cleared
30 up. So dissolution under vacuum or removal of foams intermittently during dissolution leads
to faster dissolution and may not require the lowering of the temperature of the mixture.
18
In one aspect, the method discloses making a stable nanomicellar ophthalmic
formulation comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
6.9, 7.8, 9.4 and 15.9.
In another aspect, the method of making a stable nanomicellar ophthalmic
5 formulation comprises an amorphous form of cyclosporine.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty
acid, a polyalkoxylated alcohol, and an aqueous vehicle, wherein the ophthalmic
formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with a polyoxyl lipid or fatty acid at a temperature
of 127-130°C until completely dissolved to form a mixture A;
b) adding the polyalkoxylated alcohol to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
15 In one aspect, the method of making a stable nanomicellar ophthalmic formulation
comprises a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9.
In another aspect, the method of making a stable nanomicellar ophthalmic
formulation comprises an amorphous form of cyclosporine.
20 In another aspect, the method of making a stable nanomicellar ophthalmic
formulation comprises cyclosporine in a form having characteristic XRD peaks at 2-theta
(deg.) 7.4, 8.7, 14.4 and 17.5.
In yet another aspect, the method of making a stable nanomicellar ophthalmic
formulation comprises cyclosporine in a form with characteristic XRD peaks at 2-theta
25 (deg.) 8.5, 9.3, 11.6 and 20.3.
In yet another aspect, the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty
acid, a polyalkoxylated alcohol, and an aqueous vehicle, wherein the ophthalmic
formulation is made by a method comprising the steps of:
30 a) mixing the polyoxyl lipid or fatty acid and polyalkoxylated alcohol at a
temperature of 127-130°C to form a mixture A;
19
b) adding cyclosporine to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
In one aspect, the method of making a stable nanomicellar ophthalmic formulation
5 comprises a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9.
In another aspect, the method of making a stable nanomicellar ophthalmic
formulation comprises an amorphous form of cyclosporine.
In another aspect, the method of making a stable nanomicellar ophthalmic
10 formulation comprises a cyclosporine in a form having characteristic XRD peaks at 2-theta
(deg.) 7.4, 8.7, 14.4 and 17.5.
In yet another aspect, the method of making a stable nanomicellar ophthalmic
formulation comprises cyclosporine in a form with characteristic XRD peaks at 2-theta
(deg.) 8.5, 9.3, 11.6 and 20.3.
15 Another embodiment of the present disclosure is a stable nanomicellar ophthalmic
formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated
alcohol, wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the polyoxyl lipid or fatty acid at a temperature
of 55°C or above to form a mixture A; and
20 b) lowering the temperature of mixture A to a temperature not higher than 40°C
prior to the complete dissolution of the cyclosporine.
Preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is done at a
temperature of 55°C-60°C to form a mixture A. Preferably, mixture A is lowered to a
temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
25 preferably, mixing the cyclosporine with the polyoxyl lipid is done at a temperature of
55°C±2°C to form a mixture A; and mixture A is lowered to a temperature not higher than
35°C±2°C prior to the complete dissolution of the cyclosporine. Further, the method
comprises adding polyalkoxylated alcohol and then mixing the resulting mixture with an
aqueous vehicle at 35°C±2°C. In another aspect, the resulting mixture is mixed with the
30 aqueous vehicle at a temperature of 55±2°C.
20
In one aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
6.9, 7.8, 9.4 and 15.9.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
5 formulation comprising an amorphous form of cyclosporine.
In another aspect, the present invention provides a stable nanomicellar ophthalmic
formulation comprising: cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated
alcohol, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the polyoxyl lipid or fatty acid at a temperature
of 55°C±2°C or above to form a mixture A;
b) keeping mixture A under vacuum to remove foam;
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine, adding
15 polyalkoxylated alcohol; and
d) mixing the resulting mixture with the aqueous vehicle.
In one aspect, the aqueous vehicle is mixed at a temperature of 35°C±2°C. In
another aspect, the aqueous vehicle is mixed at a temperature of 55±2°C.
In one aspect, the stable nanomicellar ophthalmic formulation compnses a
20 cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9.
In another aspect, the stable nanomicellar ophthalmic formulation comprises an
amorphous form of cyclosporine.
In yet another aspect, the present disclosure provides a stable nanomicellar
ophthalmic formulation comprising:
25 cyclosporine, a polyoxyl lipid or fatty acid, a polyalkoxylated alcohol, and an aqueous
vehicle wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with a polyoxyl lipid or fatty acid at a temperature
of 127-130°C until completely dissolved to form a mixture A;
b) adding the polyalkoxylated alcohol to mixture A at 127-130°C; and
21
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
In one aspect, the stable nanomicellar ophthalmic formulation compnses a
cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9.
5 In another aspect, the stable nanomicellar ophthalmic formulation comprises an
amorphous form of cyclosporine.
In another aspect, the stable nanomicellar ophthalmic formulation comprises a
cyclosporine in a form having characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7, 14.4 and
17.5.
10 In yet another aspect, the stable nanomicellar ophthalmic formulation comprises
cyclosporine in a form with characteristic XRD peaks at 2-theta (deg.) 8.5, 9.3, 11.6 and
20.3.
In yet another aspect, the present disclosure provides a stable nanomicellar ophthalmic
formulation comprising:
15 cyclosporine, a polyoxyl lipid or fatty acid, a polyalkoxylated alcohol, and an aqueous
vehicle wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the polyoxyl lipid or fatty acid and polyalkoxylated alcohol at a
temperature of 127-130°C to form a mixture A;
b) adding cyclosporine to mixture A at 127-130°C; and
20 c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
In one aspect, the stable nanomicellar ophthalmic formulation compnses a
cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9.
In another aspect, the stable nanomicellar ophthalmic formulation comprises an
25 amorphous form of cyclosporine.
In another aspect, the stable nanomicellar ophthalmic formulation compnses
cyclosporine in a form having characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7, 14.4 and
17.5.
22
In yet another aspect, the stable nanomicellar ophthalmic formulation comprises
cyclosporine in a form with characteristic XRD peaks at 2-theta (deg.) 8.5, 9.3, 11.6 and
20.3.
Another embodiment of the present disclosure, is a method of making a stable
5 nanomicellar ophthalmic formulation comprising a cyclosporine, a polyoxyl lipid or fatty
acid, and a polyalkoxylated alcohol, wherein the ophthalmic formulation is made by a
method comprising the steps of: mixing the cyclosporine with the polyoxyl lipid or fatty
acid at a temperature of 55°C or above to form a mixture A; and preventing the formation
of a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7, 14.4 and
10 17.5 (B in Figure 2) by altering the temperature of mixture A prior to the complete
dissolution of the cyclosporine.
Preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is done at a
temperature of 55°C-60°C to form a mixture A. Preferably, mixture A is lowered to a
temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
15 preferably, mixing the cyclosporine with the polyoxyl lipid is done at a temperature of
55°C±2°C to form a mixture A; and mixture A is lowered to a temperature not higher than
35°C±2°C prior to the complete dissolution of the cyclosporine. In one aspect, the method
of making a stable nanomicellar ophthalmic formulation comprises cyclosporine with
characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9 (A in Figure 2). In another
20 aspect, the method of making a stable nanomicellar ophthalmic formulation comprises an
amorphous form of cyclosporine. Further, the method comprises adding the polyalkoxylated
alcohol and then mixing the resulting mixture with an aqueous vehicle at 35°C±2°C. In
another aspect, the resulting mixture is mixed with the aqueous vehicle at a temperature of
55±2°C.
25 Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty
acid, and a polyalkoxylated alcohol, wherein the ophthalmic formulation is made by a
method comprising the steps of: mixing the cyclosporine with the polyoxyl lipid or fatty
acid at a temperature of 55°C or above to form a mixture A; and applying vacuum to the
30 mixture to prevent the formation of a cyclosporine form with characteristic XRD peaks at
2-theta (deg.) 7.4, 8.7, 14.4 and 17.5 (Bin Figure 2).
23
Preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is done at a
temperature of 55°C-60°C to form a mixture A. Optionally, the mixture A is lowered to a
temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with the polyoxyl lipid is done at a temperature of
5 55°C±2°C to form a mixture A; optionally lowering to a temperature not higher than
35°C±2°C prior to the complete dissolution of the cyclosporine. Further, the method
comprises adding the polyalkoxylated alcohol and then mixing the resulting mixture with
an aqueous vehicle at 35°C±2°C. In another aspect, the resulting mixture is mixed with the
aqueous vehicle at a temperature of 55±2°C.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty
acid, and a polyalkoxylated alcohol, wherein the ophthalmic formulation is made by a
method comprising the steps of: mixing the cyclosporine with the polyoxyl lipid or fatty
acid at a temperature of 55°C or above to form a mixture A; and preventing the formation
15 of a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 8.5, 9.3, 11.6 and
20.3 (C in Figure 2) by altering the temperature of mixture A prior to the complete
dissolution of the cyclosporine.
Preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is done at a
temperature of 55°C-60°C to form a mixture A. Further, mixture A is lowered to a
20 temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with the polyoxyl lipid is done at a temperature of
55°C±2°C to form a mixture A; and mixture A is then lowered to a temperature not higher
than 35°C±2°C prior to the complete dissolution of the cyclosporine. In one aspect, the
method of making a stable nanomicellar ophthalmic formulation comprises cyclosporine
25 with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9 (A in Figure 2). In
another aspect, the method of making a stable nanomicellar ophthalmic formulation
comprises an amorphous form of cyclosporine. Further, the method comprises adding the
polyalkoxylated alcohol and then mixing the resulting mixture with an aqueous vehicle at
35°C±2°C. In another aspect, the resulting mixture is mixed with the aqueous vehicle at a
30 temperature of 55±2°C.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty
acid, and a polyalkoxylated alcohol, wherein the ophthalmic formulation is made by a
24
method comprising the steps of: mixing the cyclosporine with the polyoxyl lipid or fatty
acid at a temperature of 55°C or above to form a mixture A; and applying vacuum to the
mixture to prevent the formation of a cyclosporine form with characteristic XRD peaks at
2-theta (deg.) 8.5, 9.3, 11.6 and 20.3 (C in Figure 2).
5 Preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is done at a
temperature of 55°C-60°C to form a mixture A. Optionally, the mixture A is lowered to a
temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with the polyoxyl lipid is done at a temperature of
55°C±2°C to form a mixture A; optionally lowering to a temperature not higher than
10 35°C±2°C prior to the complete dissolution of the cyclosporine. Further, the method
comprises adding the polyalkoxylated alcohol and then mixing the resulting mixture with
an aqueous vehicle at 35°C±2°C. In another aspect, the resulting mixture is mixed with the
aqueous vehicle at a temperature of 55±2°C.
Another embodiment of the present disclosure is a stable nanomicellar ophthalmic
15 formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated
alcohol, wherein the formulation is a solution; and the formulation exhibits stability at room
temperature (20-25°C) for 6 to at least 24 months. Typically, the ophthalmic formulations
are stable when maintained at room temperature for at least 6 months, at least 8 months, at
least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months,
20 at least 20 months and at least 24 months.
In another embodiment, the present disclosure provides a stable nanomicellar
ophthalmic formulation that exhibits stability at 2°C to 8°C for 6 to at least 24 months.
Typically, the ophthalmic formulations are stable when maintained at 2°C to 8°C for at least
6 months, at least 8 months, at least 10 months, at least 12 months, at least 14 months, at
25 least 16 months, at least 18 months, at least 20 months and at least 24 months.
Another embodiment of the present disclosure is a method of treating or preventing
an ocular disease or condition, the method comprising administering the formulation of any
of the preceding embodiments after 6 to at least 24 months of the manufacture of the
formulation to a patient in need thereof.
30 In another embodiment, the present disclosure is a stable nanomicellar ophthalmic
formulation of any of the preceding embodiments, for use in the treatment of an ocular
disease or condition. Preferably, the ocular disease or condition is dry eye syndrome.
25
Materials useful in the formulations of the present disclosure include, but are not
limited to, those disclosed in U.S. Patent No. 10,918,694.
As they are used here, the terms "cyclosporin", "cyclosporine", "cyclosporine A",
or "CsA" may be used interchangeably and includes pharmaceutically acceptable salts of
5 the same.
As used herein in connection with numerical values, the terms "approximately" and
"about" mean +/-10% of the indicated value, including the indicated value.
As used herein, the term "substantially free" refers to an amount of 10% or less of
the indicated substance, such as another form, preferably 8%, 5%, 4%, 3%, 2%, 1%, 0.5%,
10 or less of another form.
As used herein, the term "polyoxyl lipid or fatty acid" refers to mono- and diesters
of lipids or fatty acids and polyoxyethylene diols. Polyoxyl lipids or fatty acids may be
numbered ("n") according to the average polymer length of the oxyethylene units (e.g., 40,
60, 80, 100) as is well understood in the art. The term "n 40 polyoxyl lipid" means that the
15 polyoxyl lipid or fatty acid has an average oxyethylene polymer length equal to or greater
than 40 units. Stearate hydrogenated castor oil and castor oil are common lipids/fatty acids
commercially available as polyoxyl lipids or fatty acid, however, it is understood that any
lipid or fatty acid could polyoxylated to become a polyoxyl lipid or fatty acid as
contemplated herein. Examples of polyoxyl lipid or fatty acids include without limitation
20 hydrogenated polyoxyl castor oil such as HCO-40, HCO-60, HCO-80, HCO-100, or
polyoxyl 40 stearate, polyoxyl 35 castor oil.
As used herein, the term "micelle" or "nanomicelle" refers to an aggregate (or
cluster) of surfactant molecules. Micelles only form when the concentration of surfactant is
greater than the critical micelle concentration (CMC). Surfactants are chemicals that are
25 amphipathic, which means that they contain both hydrophobic and hydrophilic groups.
Micelles can exist in different shapes, including spherical, cylindrical, and discoidal. A
micelle comprising at least two different molecular species is a mixed micelle. The in some
embodiments, ophthalmic compositions of the present disclosure include an aqueous, clear,
mixed micellar solution.
30 In some embodiments the formulations include, but are not limited to, nanomicelles,
as disclosed in U.S. Patent No. 10,918,694. For example, ophthalmic compositions can be
administered topically to the eye as biocompatible, aqueous, clear mixed micellar solutions.
26
The compositions have the drugs incorporated and/or encapsulated in micelles which are
dispersed in an aqueous medium.
In some aspects of embodiments, the polyoxyl lipid or fatty acid is a polyoxyl castor
oil. In some embodiments, the polyoxyl lipid or fatty acid is one or more selected from
5 hydrogenated polyoxyl castor oil such as HCO-40, HCO-60, HCO-80 or HCO-100. In some
embodiments the polyoxyl lipid or fatty acid (such as a polyoxyl castor oil such as HCO-
60, HCO-80 or HCO-100) is present between 0.5 and 2%, or 0.7 and 2%, or 1 and 6%; or 2
and 6%; or 2 and 6%; or 3 and 6%; or 4 and 6%; or 2 and 5%; or 3 and 5%; or 3 and 5%; or
2 and 6%; or about 4%; or greater than 0.7%; or greater than 1%, or greater than 1.5%; or
10 greater than 2%; or greater than 3%; or greater than 4% by weight of the formulation. In
some embodiments, the polyoxyl lipid is HCO-40. In some aspects of embodiments, the
polyoxyl lipid is HCO-60. In some embodiments, the polyoxyl lipid is HCO-80. In some
embodiments, the polyoxyl lipid is HCO-100.
In some aspects of embodiments, the formulation includes a polyalkoxylated
15 alcohol. In some embodiments, the polyalkoxylated alcohol is octoxynol-40. In some
aspects of embodiments, the formulation includes a polyalkoxylated alcohol (such as
octoxynol-40) present between 0.002 and 4%; or between 0.005 and 3%; or between 0.005
and 2%; or between 0.005 and 1%; or between 0.005 and 0.5%; or between 0.005 and 0.1%;
or between 0.005 and 0.05%; or between 0.008 and 0.02%; or between 0.01 and 0.1%; or
20 between 0.02 and 0.08%; or between 0.005 and 0.08%; or about 0.05%, or about 0.01% by
weight of the formulation.
One embodiment of the present disclosure is a stable nanomicellar ophthalmic
formulation. The ophthalmic formulation comprises cyclosporine, a hydrogenated polyoxyl
castor oil, and octoxynol-40. Preferably, the hydrogenated polyoxyl castor oil is
25 hydrogenated 40 polyoxyl castor oil (HCO-40). Preferably, the ophthalmic formulation
comprises 0.09 wt% of cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and
0.05 wt% of octoxynol-40.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising cyclosporine, a hydrogenated polyoxyl
30 castor oil, and octoxynol-40, wherein the ophthalmic formulation is made by a method
comprising the steps of:
27
a) m1xmg the cyclosporine with a hydrogenated polyoxyl castor oil at a
temperature of 55°C or above to form a mixture A; and
b) lowering the temperature of mixture A to a temperature not higher than 40°C
prior to the complete dissolution of the cyclosporine.
5 Preferably, mixing the cyclosporine with a hydrogenated polyoxyl castor oil is done
at a temperature of 55°C-60°C to form a mixture A. Preferably, mixture A is lowered to a
temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is done at a
temperature of 55°C±2°C to form a mixture A; and mixture A is then lowered to a
10 temperature not higher than 35°C±2°C prior to the complete dissolution of the cyclosporine.
Further, the method comprises adding octoxynol-40 and then mixing the resulting mixture
with an aqueous vehicle at 35°C±2°C. In another aspect, the aqueous vehicle is mixed at a
temperature of 55±2°C.
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor
15 oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
40.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising cyclosporine, a hydrogenated polyoxyl
20 castor oil, and octoxynol-40, wherein the ophthalmic formulation is made by a method
comprising the steps of:
a) mixing the cyclosporine with a hydrogenated polyoxyl castor oil at a
temperature of 55°C or above to form a mixture A; and
b) keeping mixture A under vacuum to remove foam;
25 c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine, adding octoxynol-
40; and
d) mixing the resulting mixture with the aqueous vehicle.
Preferably, mixing the cyclosporine with a hydrogenated polyoxyl castor oil is done
30 at a temperature of 55°C-60°C to form a mixture A. Optionally, mixture A is lowered to a
temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
28
preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is done at a
temperature of 55°C±2°C to form a mixture A; and optionally lowering the temperature to
not higher than 35°C±2°C prior to the complete dissolution of the cyclosporine. Further, the
method comprises adding octoxynol-40 and then mixing the resulting mixture with an
5 aqueous vehicle at 35°C±2°C. In another aspect, the aqueous vehicle is mixed at a
temperature of 55±2°C.
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor
oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
10 40.
Another embodiment of the present disclosure is a stable nanomicellar ophthalmic
formulation comprising cyclosporine, a hydrogenated polyoxyl castor oil, and octoxynol-
40, wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated polyoxyl castor oil at a
15 temperature of 55°C or above to form a mixture A; and
b) lowering the temperature of mixture A to a temperature not higher than 40°C
prior to the complete dissolution of the cyclosporine.
Preferably, mixing the cyclosporine with a hydrogenated polyoxyl castor oil is done
at a temperature of 55°C-60°C to form a mixture A. Preferably, mixture A is lowered to a
20 temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is done at a
temperature of 55°C±2°C to form a mixture A; and then mixture A is lowered to a
temperature not higher than 35°C±2°C prior to the complete dissolution of the cyclosporine.
Further, the method comprises adding octoxynol-40 and then mixing the resulting mixture
25 with an aqueous vehicle at 35°C±2°C. In another aspect, the aqueous vehicle is mixed at a
temperature of 55±2°C.
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor
oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
30 40.
29
Another embodiment of the present disclosure relates to a method of making a stable
nanomicellar ophthalmic formulation comprising:
cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated alcohol, wherein the
ophthalmic formulation is made by a method comprising the steps of:
5 a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine;
c) adding octoxynol-40; and
d) mixing the resulting mixture with the aqueous vehicle.
Another embodiment of the present disclosure is a stable nanomicellar ophthalmic
formulation comprising cyclosporine, a hydrogenated polyoxyl castor oil, and octoxynol-
40, wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with a hydrogenated polyoxyl castor oil at a
15 temperature of 55°C or above to form a mixture A; and
b) keeping mixture A under vacuum to remove foam;
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine;
d) adding octoxynol-40; and
20 e) mixing the resulting mixture with the aqueous vehicle.
Preferably, mixing the cyclosporine with a hydrogenated polyoxyl castor oil is done
at a temperature of 55°C-60°C to form a mixture A. Optionally, mixture A is lowered to a
temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is done at a
25 temperature of 55°C±2°C to form a mixture A; and optionally lowering the temperature to
not higher than 35°C±2°C prior to the complete dissolution of the cyclosporine. Further, the
method comprises adding octoxynol-40 and then mixing the resulting mixture with an
aqueous vehicle at 35°C±2°C. In another aspect, the aqueous vehicle is mixed at a
temperature of 55±2°C.
30
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor
oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
40.
5 Another embodiment of the present disclosure is a stable nanomicellar ophthalmic
formulation comprising: cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40,
and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine;
c) adding octoxynol-40; and
d) mixing the resulting mixture with the aqueous vehicle.
15 In one aspect, mixing the aqueous vehicle to a temperature of at 35°C±2°C. In
another aspect, mixing the aqueous vehicle to a temperature of 55±2°C.
In one embodiment, the present invention provides a stable nanomicellar ophthalmic
formulation comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
6.9, 7.8, 9.4 and 15.9.
20 In another embodiment, the present invention provides a stable nanomicellar
ophthalmic formulation comprising an amorphous form of cyclosporine.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising cyclosporine, a hydrogenated polyoxyl
castor oil, and octoxynol-40, wherein the ophthalmic formulation is made by a method
25 comprising the steps of: mixing the cyclosporine with a hydrogenated polyoxyl castor oil at
a temperature of 55°C or above to form a mixture A; and preventing the formation of a
cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7, 14.4 and 17.5
(Bin Figure 2) by altering the temperature of mixture A prior to the complete dissolution of
the cyclosporine.
31
Preferably, mixing the cyclosporine with a hydrogenated polyoxyl castor oil is done
at a temperature of 55°C-60°C to form a mixture A. Further, mixture A is lowered to a
temperature of 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is done at a
5 temperature of 55°C±2°C to form a mixture A; and then mixture A is lowered to a
temperature not higher than 35°C±2°C prior to the complete dissolution of the cyclosporine.
In one aspect, the stable nanomicellar ophthalmic formulation comprises cyclosporine with
characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9 (A in Fig.2). In another
aspect, the stable nanomicellar ophthalmic formulation comprises an amorphous form of
10 cyclosporine. Further, the method comprises adding the octoxynol-40 and then mixing the
resulting mixture with an aqueous vehicle at 35°C±2°C. In another aspect, the resulting
mixture is mixed with the aqueous vehicle at a temperature of 55±2°C. Preferably, the
hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More
preferably, the ophthalmic formulation comprises 0.09 wt% of cyclosporine, 1.0 wt% of
15 hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-40.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising cyclosporine, a hydrogenated polyoxyl
castor oil, and octoxynol-40, wherein the ophthalmic formulation is made by a method
comprising the steps of: mixing the cyclosporine with the hydrogenated polyoxyl castor oil
20 at a temperature of 55°C or above to form a mixture A; and preventing the formation of a
cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 8.5, 9.3, 11.6 and 20.3
(C in Figure 2) by altering the temperature of mixture A prior to the complete dissolution of
the cyclosporine.
Preferably, mixing the cyclosporine with a hydrogenated polyoxyl castor oil is done
25 at a temperature of 55°C-60°C to form a mixture A. Further, mixture A is then lowered to
a temperature of 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is done at a
temperature of 55°C±2°C to form a mixture A; and then the temperature of mixture A is
lowered to a temperature not higher than 35°C±2°C prior to the complete dissolution of the
30 cyclosporine In one aspect, the method of making a stable nanomicellar ophthalmic
formulation comprises cyclosporine with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8,
9.4 and 15.9 (A in Figure 2). In another aspect, the method of making a stable nanomicellar
ophthalmic formulation comprises an amorphous form of cyclosporine. Further, the method
32
comprises adding octoxynol-40 and then mixing the resulting mixture with an aqueous
vehicle at 35°C±2°C. In another aspect, the aqueous vehicle is mixed at a temperature of
55±2°C. Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl
castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt% of
5 cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
40.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising cyclosporine, a hydrogenated polyoxyl
castor oil, and octoxynol-40, wherein the ophthalmic formulation is made by a method
10 comprising the steps of: mixing the cyclosporine with hydrogenated polyoxyl castor oil at a
temperature of 55°C or above to form a mixture A; and applying vacuum to the mixture to
prevent the formation of a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
7.4, 8.7, 14.4 and 17.5 (Bin Figure 2).
Preferably, mixing the cyclosporine with hydrogenated polyoxyl castor oil is done
15 at a temperature of 55°C-60°C to form a mixture A. Optionally, the mixture A is lowered
to a temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is done at a
temperature of 55°C±2°C to form a mixture A; optionally lowering to a temperature not
higher than 35°C±2°C prior to the complete dissolution of the cyclosporine. Further, the
20 method comprises adding the octoxynol-40 and then mixing the resulting mixture with an
aqueous vehicle at 35°C±2°C. In another aspect, the resulting mixture is mixed with the
aqueous vehicle at a temperature of 55±2°C. Preferably, cyclosporine is present in a
cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9 (A
in Figure 2). In another embodiment, preferably, cyclosporine is present in amorphous form.
25 Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil
(HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
40.
Another embodiment of the present disclosure is a method of making a stable
30 nanomicellar ophthalmic formulation comprising cyclosporine, a hydrogenated polyoxyl
castor oil, and octoxynol-40, wherein the ophthalmic formulation is made by a method
comprising the steps of: mixing the cyclosporine with hydrogenated polyoxyl castor oil at a
temperature of 55°C or above to form a mixture A; and applying vacuum to the mixture to
33
prevent the formation of a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
8.5, 9.3, 11.6 and 20.3 (C in Figure 2).
Preferably, mixing the cyclosporine with hydrogenated polyoxyl castor oil is done
at a temperature of 55°C-60°C to form a mixture A. Optionally, the mixture A is lowered
5 to a temperature 35°C- 40°C prior to the complete dissolution of the cyclosporine. More
preferably, mixing the cyclosporine with hydrogenated polyoxyl castor oil is done at a
temperature of 55°C±2°C to form a mixture A; optionally lowering to a temperature not
higher than 35°C±2°C prior to the complete dissolution of the cyclosporine. Further, the
method comprises adding the octoxynol-40 and then mixing the resulting mixture with an
10 aqueous vehicle at 35°C±2°C. In another aspect, the resulting mixture is mixed with the
aqueous vehicle at a temperature of 55±2°C. Preferably, cyclosporine is present in a
cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9 (A
in Figure 2). In another embodiment, preferably, cyclosporine is present in amorphous form.
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil
15 (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
40.
In yet another aspect, the present disclosure 1s a method of making a stable
nanomicellar ophthalmic formulation comprising:
20 cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
25 c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
In one aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising a cyclosporine form with characteristic XRD peaks at
2-theta (deg.) 6.9, 7.8, 9.4 and 15.9.
30 In another aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising an amorphous form of cyclosporine.
34
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising:
cyclosporine, hydrogenated polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the method comprising the steps of:
5 a) mixing the cyclosporine with the hydrogenated polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C;
10 wherein cyclosporine is present in a form having characteristic XRD peaks at 2-theta (deg.)
7.4, 8.7, 14.4 and 17.5 (Bin Figure 2).
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor
oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
15 40.
Another embodiment of the present disclosure is a method of making a stable
nanomicellar ophthalmic formulation comprising:
cyclosporine, hydrogenated polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the method comprising the steps of:
20 a) mixing the cyclosporine with the hydrogenated polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of
127-130°C,
25 wherein cyclosporine is present in a form with characteristic XRD peaks at 2-theta (deg.)
8.5, 9.3, 11.6 and 20.3 (C in Figure 2).
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor
oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
30 40.
WO 2023/021492 PCT/IB2022/057851
34
35
In yet another aspect, the present disclosure 1s a method of making a stable
nanomicellar ophthalmic formulation comprising:
cyclosporine, hydrogenated polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the method comprising the steps of:
5 a) mixing the hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a
temperature of 127-130°C to form a mixture A;
b) adding cyclosporine to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
In one aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising a cyclosporine form with characteristic XRD peaks at
2-theta (deg.) 6.9, 7.8, 9.4 and 15.9.
In another aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising an amorphous form of cyclosporine.
15 In another aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks
at 2-theta (deg.) 7.4, 8.7, 14.4 and 17.5.
In another aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks
20 at 2-theta (deg.) 8.5, 9.3, 11.6 and 20.3.
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor
oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
40.
25 In yet another aspect, the present disclosure is a stable nanomicellar ophthalmic
formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
36
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
5 127-130°C.
In one aspect, the present disclosure a stable nanomicellar ophthalmic formulation
comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9.
In another aspect, the present disclosure is a stable nanomicellar ophthalmic
10 formulation comprising an amorphous form of cyclosporine.
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor
oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
40.
15 In yet another embodiment of the present disclosure 1s a stable nanomicellar
ophthalmic formulation comprising:
cyclosporine, hydrogenated polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated polyoxyl castor oil at a
20 temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C;
wherein cyclosporine is present in a form having characteristic XRD peaks at 2-theta (deg.)
25 7.4, 8.7, 14.4 and 17.5 (Bin Figure 2).
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor
oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
40.
37
Yet another embodiment of the present disclosure 1s a stable nanomicellar
ophthalmic formulation comprising:
cyclosporine, hydrogenated polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
5 a) mixing the cyclosporine with the hydrogenated polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of
127-130°C,
10 wherein cyclosporine is present in a form with characteristic XRD peaks at 2-theta (deg.)
8.5, 9.3, 11.6 and 20.3 (C in Figure 2).
Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor
oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09 wt % of
cyclosporine, 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-
15 40.
In yet another aspect, the present disclosure is a stable nanomicellar ophthalmic
formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
20 a) mixing the hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a
temperature of 127-130°C to form a mixture A;
b) adding cyclosporine to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
25 In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation
comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9.
In another aspect, the present disclosure is a stable nanomicellar ophthalmic
formulation comprising an amorphous form of cyclosporine.
38
In another aspect, the present disclosure is a stable nanomicellar ophthalmic
formulation comprising cyclosporine in a form having characteristic XRD peaks at 2-theta
(deg.) 7.4, 8.7, 14.4 and 17.5.
In another aspect, the present disclosure is a stable nanomicellar ophthalmic
5 formulation comprising cyclosporine in a form having characteristic XRD peaks at 2-theta
(deg.) 8.5, 9.3, 11.6 and 20.3.
Another embodiment of the present disclosure is a stable nanomicellar ophthalmic
formulation comprising cyclosporine, a hydrogenated 40 polyoxyl castor oil (HCO-40), and
octoxynol-40, wherein the formulation is a solution; and the formulation exhibits stability
10 at room temperature (20-25°C) for 6 to at least 24 months. Typically, the ophthalmic
formulations are stable when maintained at room temperature for at least 6 months, at least
8 months, at least 10 months, at least 12 months, at least 14 months, at least 16 months, at
least 18 months, at least 20 months and at least 24 months. More preferably, the stable
nanomicellar ophthalmic formulation is a solution and comprises 0.09 wt% of cyclosporine,
15 1.0 wt% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-40.
Another embodiment of the present disclosure is a method of treating or preventing
an ocular disease or condition, for example, dry eye, the method comprising administering
a stable nanomicellar ophthalmic formulation comprising 0.09 wt% of cyclosporine, 1.0 wt
% of hydrogenated 40 polyoxyl castor oil, and 0.05 wt% of octoxynol-40 after 6 to at least
20 24 months of the manufacture of the formulation to a patient in need thereof.
The cyclosporine present in certain formulations according to embodiments of this
disclosure is preferably amorphous when it is in solution. Alternatively, the cyclosporine
may be present as a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9,
7.8, 9.4 and 15.9 in solution. In certain embodiments, there is no crystalline cyclosporine
25 present in a solution according to the present disclosure. In certain embodiments, the
formulations are substantially free of a cyclosporine form with characteristic XRD peaks at
2-theta (deg.) 7.4, 8.7, 14.4 and 17.5. In alternative embodiments, the formulations
according to this disclosure are substantially free of a cyclosporine form with characteristic
XRD peaks at 2-theta (deg.) 8.5, 9.3, 11.6 and 20.3.
30 Additional Formulation Ingredients
The compositions of the present disclosure may also contain other components such
as, but not limited to, additives, adjuvants, buffers, tonicity agents, bioadhesive polymers,
39
and preservatives. In any of the compositions of this disclosure for topical to the eye, the
mixtures are preferably formulated at about pH 5 to about pH 8. This pH range may be
achieved by the addition of buffers to the composition as described in the examples. In an
embodiment, the pH range in the composition in a formulation is about pH 6.5 to about pH
5 7.2. It should be appreciated that the compositions of the present disclosure may be buffered
by any common buffer system such as phosphate, borate, acetate, citrate, carbonate and
borate-polyol complexes, with the pH and osmolality adjusted in accordance with well-
known techniques to proper physiological values. The mixed micellar compositions of the
present disclosure are stable in buffered aqueous solution. That is, there is no adverse
10 interaction between the buffer and any other component that would cause the compositions
to be unstable.
Tonicity agents include, for example, mannitol, sodium chloride, sodium nitrate,
sodium sulfate, dextrose, xylitol or combinations thereof. These tonicity agents may be used
to adjust the osmolality of the compositions. In an aspect, the osmolality of the formulation
15 is adjusted to be in the range of about 150 to about 200 mOsmol/kg. In a preferred aspect,
the osmolality of the formulation is adjusted to between about 160 to about 190 mOsmol/kg.
An additive such as a sugar, a glycerol, and other sugar alcohols, can be included in
the compositions of the present disclosure. Pharmaceutical additives can be added to
increase the efficacy or potency of other ingredients in the composition. For example, a
20 pharmaceutical additive can be added to a composition of the present disclosure to improve
the stability of the calcineurin inhibitor, to adjust the osmolality of the composition, to adjust
the viscosity of the composition, or for another reason, such as effecting drug delivery. Non-
limiting examples of pharmaceutical additives of the present disclosure include sugars, such
as, trehalose, mannose, D-galactose, and lactose. In an embodiment, the sugars can be
25 incorporated into a composition prior to hydrating the thin film (i.e. internally). In another
embodiment, the sugars can be incorporated into a composition during the hydration step
(i.e. externally). In an embodiment, an aqueous, clear, mixed micellar solution of the present
disclosure includes additives such as sugars.
In an embodiment, compositions of the present disclosure further comprise one or
30 more bioadhesive polymers. Bioadhesion refers to the ability of certain synthetic and
biological macromolecules and hydrocolloids to adhere to biological tissues. Bioadhesion
is a complex phenomenon, depending in part upon the properties of polymers, biological
tissue, and the surrounding environment. Several factors have been found to contribute to a
40
polymer's bioadhesive capacity: the presence of functional groups able to form hydrogen
bridges (--OH, COOH), the presence and strength of anionic charges, sufficient elasticity
for the polymeric chains to interpenetrate the mucous layer, and high molecular weight.
Bioadhesion systems have been used in dentistry, orthopedics, ophthalmology, and in
5 surgical applications. However, there has recently emerged significant interest in the use of
bioadhesive materials in other areas such as soft tissue-based artificial replacements, and
controlled release systems for local release of bioactive agents. Such applications include
systems for release of drugs in the buccal or nasal cavity, and for intestinal or rectal
administration.
In an embodiment, a composition of the present disclosure includes at least one
bioadhesive polymer. The bioadhesive polymer can enhance the viscosity of the
composition and thereby increase residence time in the eye. Bioadhesive polymers of the
present disclosure include, for example, carboxylic polymers like Carbopol® (carbomers),
Noveon® (polycarbophils), cellulose derivatives including alkyl and hydroxyalkyl cellulose
15 like methylcellulose, hydroxypropylcellulose, carboxymethylcellulose, gums like locust
beam, xanthan, agarose, karaya, guar, and other polymers including but not limited to
polyvinyl alcohol, povidone, polyethylene glycol, Pluronic® (Poloxamers), tragacanth, and
hyaluronic acid; phase-transition polymers for providing sustained and controlled delivery
of enclosed medicaments to the eye (e.g., alginic acid, carrageenans (e.g., Eucheuma),
20 xanthan and locust bean gum mixtures, pectins, cellulose acetate phthalate,
alkylhydroxyalkyl cellulose and derivatives thereof, hydroxyalkylated polyacrylic acids and
derivatives thereof, poloxamers and their derivatives, etc. Physical characteristics in these
polymers can be mediated by changes in environmental factors such as ionic strength, pH,
or temperature alone or in combination with other factors. In an embodiment, the optional
25 one or more bioadhesive polymers is present in the composition from about 0.01 wt% to
about 10 wt %/volume, preferably from about 0.1 to about 5 wt %/volume. In an
embodiment, the compositions of the present disclosure further comprise at least one
hydrophilic polymer excipient selected from, for example, PVP-K-30, PVP-K-90, HPMC,
HEC, and polycarbophil. In an embodiment, the polymer excipient is selected from PVP-
30 K-90, PVP-K-30 or HPMC. In an embodiment, the polymer excipient is selected from PVP-
K-90 or PVP-K-30.
In an embodiment, if a preservative is desired, the compositions may optionally be
preserved with any of many well-known preservatives, including benzyl alcohol
41
with/without EDTA, benzalkonium chloride, chlorhexidine, Cosmocil® CQ, or Dowicil®
200. In certain embodiments, it may be desirable for a formulation as described herein to
not include any preservatives. In this regard, preservatives may in some embodiments not
be necessary or desirable in formulations included in single use containers. In other
5 embodiments, it may be advantageous to include preservatives, such as in certain
embodiments in which the formulations are included in a multiuse container.
In a preferable embodiment, the present disclosure relates to a method of making a
stable nanomicellar ophthalmic formulation comprising:
0.09 wt% of cyclosporine
about 1.0 wt% of hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% of octoxynol-40, and
an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
15 temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine, adding octoxynol-40; and
c) mixing the resulting mixture with the aqueous vehicle.
In another preferable embodiment, the present disclosure relates to a method of
20 making a stable nanomicellar ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
25 wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) keeping mixture A under vacuum to remove foam;
42
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine;
d) adding octoxynol-40; and
e) mixing the resulting mixture with the aqueous vehicle.
5 In one aspect, mixing the aqueous vehicle occurs at a temperature of 35°C±2°C. In
another aspect, mixing the aqueous vehicle is done at a temperature of 55±2°C.
The stable nanomicellar ophthalmic formulations of preceding embodiments further
compnse:
about 0.20-0.550 wt% sodium phosphate monobasic,
about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
about 0.3 wt% povidone,
sodium hydroxide/hydrochloric acid to adjust the pH, and
water for injection.
15 In another preferable embodiment, the present disclosure is a stable nanomicellar
ophthalmic formulation, wherein the pH of the formulation is about 5.0 to 8.0. More
preferably, the pH of the formulation is about 6.5 to 7.2.
Further, the present invention disclosure is a stable nanomicellar ophthalmic
formulation wherein the osmolality of the formulation is between about 150 to about 200
20 mOsmol/kg.
Further, the present invention disclosure is a stable nanomicellar ophthalmic
formulation wherein the mixed nanomicellar size and polydispersity index are determined
with Zetasizer, Malvern Instruments, N.J. In brief, approximately 1 ml of each formulation
was transferred to a cuvette and placed in the instrument. A laser beam of light was used to
25 determine the mixed nanomicellar size. Nanomicelles contemplated by the present
disclosure typically have a particle size in the range of about 1-100 nm; in some
embodiments, the particle size falls in the range of about 5-50 nm; in some embodiments,
the particle size falls in the range of about 10-40 nm; in some embodiments, the particle size
is about 13-16 nm.
43
In another embodiment, the present disclosure relates to a cyclosporine form with
characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9.
In yet another embodiment, the present disclosure relates to a stable nanomicellar
ophthalmic formulation comprising an amorphous form of cyclosporine.
5 In yet another embodiment, the present disclosure relates to a stable nanomicellar
ophthalmic formulation, wherein the formulation is substantially free of a cyclosporine form
with characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7, 14.4 and 17.5.
In yet another embodiment, the present disclosure is a stable nanomicellar
ophthalmic formulation, wherein the formulation is substantially free of a cyclosporine form
10 with characteristic XRD peaks at 2-theta (deg.) 8.5, 9.3, 11.6 and 20.3.The XRD data for
these cyclosporine forms are presented in Figure 2.
The compete dissolution time of cyclosporine depends on quantity of the
cyclosporine A to be dissolved based on the batch size. Generally dissolution time of
cyclosporine in hydrogenated 40 polyoxyl castor oil (Kolliphor® RH 40) at a ratio of 9:10
15 (cyclosporine : Kolliphor® RH 40) was found to be not less than 130 minutes. The
cyclosporine A slowly dissolved over a period of time with stirring and during this complete
dissolution period, the solution became clear. If the cyclosporine A re-precipitates during
the dissolution period, there is a chance that a slight turbid solution might have been
transferred to the aqueous phase during manufacturing of the batch. This may initiate a
20 seeding effect for crystal growth in the final formulation during storage, which may lead to
batch failure. Based on these observations, the solubility behavior of cyclosporine was
studied in hydrogenated 40 polyoxyl castor oil (Kolliphor® RH 40) at 55°C and 35°C. It was
found that the solution stability at 35°C was comparatively higher than the solution stability
at 55°C (see Example 2 in Table 4). Based on the above observation it is believed that if
25 cyclosporine was not completely dissolved in Kolliphor® RH 40 at 55°C, the temperature
can be reduced to 35°C. This is because the solution stability of cyclosporine at 35°C was
higher than that of 55°C.
In another aspect, when the cyclosporine is present in a form having characteristic
XRD peaks at 2-theta (deg.) 7.4, 8.7, 14.4 and 17.5 (B in Figure 2), the mixing of the
30 cyclosporine with the hydrogenated polyoxyl castor oil is done at a temperature of 127-
130°C until completely dissolved and octoxynol-40 is added to this to mixture at 127-130°C.
44
In another aspect, when the cyclosporine is present in a form having characteristic
XRD peaks at 2-theta (deg.) 8.5, 9.3, 11.6 and 20.3 (C in Figure 2)., the mixing of the
cyclosporine with the hydrogenated polyoxyl castor oil is done at a temperature of 127-
130°C until completely dissolved and octoxynol-40 is added to this to mixture at 127-130°C.
5 In certain aspects of the present invention, in the step of lowering the temperature of
mixture A, mixture A is lowered to a temperature of 35°C±2°C. The step of lowering the
temperature of mixture A may occur in less than 65 minutes. Preferably, at about 60 ruins.
In certain aspects, water may be added after the step of lowering the temperature of mixture
A.
10 In another aspect, the present disclosure relates to a stable nanomicellar ophthalmic
formulation, prepared by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
15 to the complete dissolution of the cyclosporine;
c) adding octoxynol-40; and
d) then mixing the resulting mixture with an aqueous vehicle at 35°C±2°C,
wherein the mixture A is lowered to a temperature of 35°C±2°C in less than 65
minutes. Preferably, the mixture A is lowered to a temperature of 35°C±2°C in 60
20 minutes.
In one aspect, mixing the mixture A of step (a) for 20-30 minutes. Preferably, 20-25
minutes, more preferably, mixing the mixture A of step (a) for 20 ±2 minutes.
In one aspect, lowering the temperature of to 35°C±2°C and stirring for 60-70
minutes. Preferably, stirring the mixture for 60±5 minutes at a temperature of 35°C±2°C.
25 In yet another aspect, the present invention relates to a method of making a stable
nanomicellar ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40,
30 about 0.20-0.550 wt% sodium phosphate monobasic,
45
about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
about 0.3 wt% povidone,
sodium hydroxide/hydrochloric acid to adjust the pH, and
5 water for injection,
wherein the method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine, adding octoxynol-40 to form an API
mixture;
c) adding the API mixture to Water for Injection (WFI);
d) adding remaining excipients to step (c) in the order of sodium phosphate
monobasic, then sodium phosphate dibasic, then sodium chloride, and then
15 polyvinylpyrrolidone;
e) adjusting the pH to 6.5 to 7.2 and bringing to the final volume with WFI.
In one aspect, the mixing cyclosporine in step (a) is done at 200-300 RPM.
In one aspect, the mixing the mixture A of step (a) is done for 20-30 minutes.
Preferably, 20-25 minutes, more preferably, 20 ±2 minutes.
20 In one aspect, the lowering the temperature of the mixture A to 35°C±2°C is done
in less than 65 minutes. Preferably, the lowering the temperature of the mixture A to
35°C±2°C is done in 40-50 minutes.
In another aspect, the mixture of step (b) is stirred for 60-70 minutes. Preferably, the
mixture is stirred for 60±5 minutes at a temperature of 35°C±2°C.
25 In one aspect, mixing the API mixture to WFI is done at a temperature of 35°C±2°C.
In another aspect, mixing the API mixture to WFI is done at a temperature of 55±2°C.
In yet another aspect, the present invention relates to a method of making a stable
nanomicellar ophthalmic formulation comprising:
0.09 wt% cyclosporine,
46
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40,
about 0.20-0.550 wt% sodium phosphate monobasic,
about 0.23-0.465 wt% sodium phosphate dibasic,
5 about 0.05 wt% sodium chloride,
about 0.3 wt% povidone,
sodium hydroxide/hydrochloric acid to adjust the pH, and
water for injection,
wherein the method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) keeping mixture A under vacuum to remove foam;
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine;
15 d) adding octoxynol-40;
e) adding the API mixture to Water for Injection (WFI);
f) adding remaining excipients to step (c) in the order of sodium phosphate
monobasic, then sodium phosphate dibasic, then sodium chloride, and then
polyvinylpyrrolidone; and
20 g) adjusting the pH to 6.5 to 7.2 and bringing to the final volume with WFI.
In one aspect, the mixing the mixture A of step (a) is done for 20-30 minutes.
Preferably, 20-25 minutes, more preferably, 20 ±2 minutes.
In one aspect, lowering the temperature of the mixture A to 35°C±2°C is done in
less than 65 minutes. Preferably, lowering the temperature of the mixture A to 35°C±2°C is
25 done in 40-50 minutes.
In one aspect, mixing the API mixture to WFI is done at a temperature of 35°C±2°C.
In another aspect, mixing the API mixture to WFI is done at a temperature of 55±2°C.
47
In a more preferred aspect, the present invention provides for a stable nanomicellar
ophthalmic formulation prepared by any of the method as described above.
In yet another aspect, the present disclosure relates to a stable nanomicellar
ophthalmic formulation comprising:
5 0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
15 In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation
comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9.
In another aspect, the present disclosure is a stable nanomicellar ophthalmic
formulation comprising an amorphous form of cyclosporine.
20 In yet another aspect, the present disclosure relates to a stable nanomicellar
ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
25 an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
48
b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C,
wherein cyclosporine is present in a form having characteristic XRD peaks at 2-theta (deg.)
5 7.4, 8.7, 14.4 and 17.5.
In yet another aspect, the present disclosure relates to a stable nanomicellar
ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
15 b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of
127-130°C,
wherein cyclosporine is present in a form with characteristic XRD peaks at 2-theta (deg.)
8.5, 9.3, 11.6 and 20.3.
20 In yet another aspect, the present disclosure is a stable nanomicellar ophthalmic
formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
25 an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a
temperature of 127-130°C to form a mixture A;
49
b) adding cyclosporine to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation
5 comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9.
In another aspect, the present disclosure is a stable nanomicellar ophthalmic
formulation comprising an amorphous form of cyclosporine.
In another aspect, the present disclosure is a stable nanomicellar ophthalmic
10 formulation comprising cyclosporine in a form having characteristic XRD peaks at 2-theta
(deg.) 7.4, 8.7, 14.4 and 17.5.
In another aspect, the present disclosure 1s a stable nanomicellar ophthalmic
formulation comprising cyclosporine.
In yet another aspect, the present disclosure relates to a stable nanomicellar
15 ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40,
about 0.20-0.550 wt% sodium phosphate monobasic,
20 about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
about 0.3 wt% povidone,
sodium hydroxide/hydrochloric acid to adjust the pH, and
water for injection,
25 wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C to form API mixture;
50
c) adding the API mixture to Water for Injection (WFI);
d) adding remaining excipients to step (c) in the order of sodium phosphate
monobasic, then sodium phosphate dibasic, then sodium chloride, and then
polyvinylpyrrolidone;
5 e) adjusting the pH to 6.5 to 7.2 and bringing to the final volume with WFI.
In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation
comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9.
In another aspect, the present disclosure is a stable nanomicellar ophthalmic
10 formulation comprising an amorphous form of cyclosporine.
In another aspect, the present disclosure is a stable nanomicellar ophthalmic
formulation comprising cyclosporine in a form having characteristic XRD peaks at 2-theta
(deg.) 7.4, 8.7, 14.4 and 17.5.
In another aspect, the present disclosure 1s a stable nanomicellar ophthalmic
15 formulation comprising cyclosporine.
In yet another aspect, the present invention relates to a method of making a stable
nanomicellar ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
20 about 0.05 wt% octoxynol-40,
about 0.20-0.550 wt% sodium phosphate monobasic,
about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
about 0.3 wt% povidone,
25 sodium hydroxide/hydrochloric acid to adjust the pH, and
water for injection,
wherein the method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
51
b) adding octoxynol-40 to mixture A at 127-130°C to form API mixture;
c) adding the API mixture to Water for Injection (WFI);
d) adding remaining excipients to step (c) in the order of sodium phosphate
monobasic, then sodium phosphate dibasic, then sodium chloride, and then
5 polyvinylpyrrolidone;
e) adjusting the pH to 6.5 to 7.2 and bringing to the final volume with WFI.
In one aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising a cyclosporine form with characteristic XRD peaks at
2-theta (deg.) 6.9, 7.8, 9.4 and 15.9.
In another aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising an amorphous form of cyclosporine.
In another aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks
at 2-theta (deg.) 7.4, 8.7, 14.4 and 17.5.
15 In another aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising cyclosporine.
In yet another aspect, the present disclosure relates to a stable nanomicellar
ophthalmic formulation comprising:
0.09 wt% cyclosporine,
20 about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40,
about 0.20-0.550 wt% sodium phosphate monobasic,
about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
25 about 0.3 wt% povidone,
sodium hydroxide/hydrochloric acid to adjust the pH, and
water for injection,
wherein the ophthalmic formulation is made by a method comprising the steps of:
52
a) mixing the hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a
temperature of 127-130°C to form a mixture A;
b) adding cyclosporin to mixture A at 127-130°C and stirred until completely
dissolved to form API mixture;
5 c) adding the API mixture to Water for Injection (WFI);
d) adding remaining excipients to step (c) in the order of sodium phosphate
monobasic, then sodium phosphate dibasic, then sodium chloride, and then
polyvinylpyrrolidone;
e) adjusting the pH to 6.5 to 7.2 and bringing to the final volume with WFI.
10 In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation
comprising a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9.
In another aspect, the present disclosure is a stable nanomicellar ophthalmic
formulation comprising an amorphous form of cyclosporine.
15 In another aspect, the present disclosure is a stable nanomicellar ophthalmic
formulation comprising cyclosporine in a form having characteristic XRD peaks at 2-theta
(deg.) 7.4, 8.7, 14.4 and 17.5.
In another aspect, the present disclosure 1s a stable nanomicellar ophthalmic
formulation comprising cyclosporine.
20 In yet another aspect, the present invention relates to a method of making a stable
nanomicellar ophthalmic formulation comprising:
0.09 wt% cyclosporine,
about 1.0 wt% hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40,
25 about 0.20-0.550 wt% sodium phosphate monobasic,
about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
about 0.3 wt% povidone,
sodium hydroxide/hydrochloric acid to adjust the pH, and
53
water for injection,
wherein the method comprising the steps of:
a) mixing the hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a
temperature of 127-130°C to form a mixture A;
5 b) adding cyclosporin to mixture A at 127-130°C and stirred until completely
dissolved to form API mixture;
c) adding the API mixture to Water for Injection (WFI);
d) adding remaining excipients to step (c) in the order of sodium phosphate
monobasic, then sodium phosphate dibasic, then sodium chloride, and then
polyvinylpyrrolidone;
e) adjusting the pH to 6.5 to 7.2 and bringing to the final volume with WFI.
In one aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising a cyclosporine form with characteristic XRD peaks at
2-theta (deg.) 6.9, 7.8, 9.4 and 15.9.
15 In another aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising an amorphous form of cyclosporine.
In another aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks
at 2-theta (deg.) 7.4, 8.7, 14.4 and 17.5.
20 In another aspect, the present disclosure is a method of making a stable nanomicellar
ophthalmic formulation comprising cyclosporine.
In one embodiment, the mixing speed, time and energy input plays a role in the
complete dissolution of cyclosporine in hydrogenated polyoxyl castor oil. When low speed
is used, time is increased for dissolution, In one aspect of the embodiment, typically
25 cyclosporine is dissolved in hydrogenated polyoxyl castor oil by stirring at approximately
100-200 RPM for 90 minutes, for 85 minutes, for 80 minutes, for 75 minutes, for 70 minutes,
for 65minutes, for 60 minutes, for 55 minutes, for 50 minutes, for 45 minutes, for 40
minutes, for 35 minutes, for 30 minutes, for 25 minutes, for 20 minutes, for 15 minutes, for
10 minutes. 200-300 RPM for 75 minutes, for 70 minutes, for 65minutes, for 60 minutes,
30 for 55 minutes, for 50 minutes, for 45 minutes, for 40 minutes, for 35 minutes, for 30
54
minutes, for 25 minutes, for 20 minutes, for 15 minutes, for 10 minutes. In another aspect,
typically cyclosporine is dissolved in hydrogenated polyoxyl castor oil by stirring at
approximately 300-400 RPM for 65minutes, for 60 minutes, for 55 minutes, for 50 minutes,
for 45 minutes, for 40 minutes, for 35 minutes, for 30 minutes, for 25 minutes, for 20
5 minutes, for 15 minutes, for 10 minutes. In another aspect, typically cyclosporine is
dissolved in hydrogenated polyoxyl castor oil by stirring at approximately 350-400 RPM
for 60 minutes, for 55 minutes, for 50 minutes, for 45 minutes, for 40 minutes, for 35
minutes, for 30 minutes, for 25 minutes, for 20 minutes, for 15 minutes, for 10 minutes. In
another aspect, typically cyclosporine is dissolved in hydrogenated polyoxyl castor oil by
10 stirring at approximately 400-450 RPM for 50 minutes, for 45 minutes, for 40 minutes, for
35 minutes, for 30 minutes, for 25 minutes, for 20 minutes, for 15 minutes, for 10 minutes.
In another aspect, typically cyclosporine is dissolved in hydrogenated polyoxyl castor oil
by stirring at approximately >450 RPM for 40 minutes, for 35 minutes, for 30 minutes, for
25 minutes, for 20 minutes, for 15 minutes, for 10 minutes, for 5 minutes. In yet another
15 aspect, cyclosporine is dissolved in hydrogenated polyoxyl castor oil by stirring at
approximately 200-300 RPM till complete dissolution.
The energy input into the mixture is defined in Equation 1 as:
EN=n3D5t/V (1)
wherein E is the theoretical energy input, n is the shear plate rpm, D is the shear plate
20 diameter, t is the time, and V is the solution volume. The energy input per volume is scale
independent. (Diaz, M., et al., "Mixing Power, External Convection, and Effectiveness in
Bioreactors," Biotechnology and Bioengineering, Vol. 51, 1996, pp. 131-140). This is a
simple, rapid and reliable way to scale up the preparation of the nanomicellar formulation.
Alternative methods of mixing, such as the use of sonicators, or the use of solvents,
25 water, or pressure, may be used to impact the temperatures or times of the respective method
steps. For example, if solvents are used, it may be possible to utilize lower temperatures in
the method steps.
In one embodiment, the mixture is mixed using a homogenizer or sonicator or mixer
comprising a stirrer. The stirrer may be located overhead or at the bottom or both overhead
30 and bottom of the homogenizer or sonicator or mixer. Preferably, the stirrer is present at
both the overhead and bottom of the homogenizer or sonicator or mixer.
55
The instant disclosure further relates to treating or preventing ocular diseases or
disorders, for example, by local administration of the formulations as described herein.
The term "treating" refers to: preventing a disease, disorder or condition from
occurring in a cell, a tissue, a system, animal or human which may be predisposed to the
5 disease, disorder and/or condition but has not yet been diagnosed as having it; stabilizing a
disease, disorder or condition, i.e., arresting its development; and/or relieving one or more
symptoms of the disease, disorder or condition, i.e., causing regression of the disease,
disorder and/or condition.
As used herein, a composition that "prevents" a disorder or condition refers to a
10 compound that, in a statistical sample, reduces the occurrence of the disorder or condition
in the treated sample relative to an untreated control sample, or delays the onset or reduces
the severity of one or more symptoms of the disorder or condition relative to the untreated
control sample.
As used herein, the term "ocular disease" refers to diseases/conditions of the eye(s)
15 that can be sight threatening, lead to eye discomfort, and may signal systemic health
problems.
A patient or subject to be treated by any of the compositions or methods of the
present disclosure can mean either a human or a non-human animal. In an embodiment, the
present disclosure provides methods for the treatment of an ocular disease in a human patient
20 in need thereof. In an embodiment, the present disclosure provides methods for the treatment
of an inflammatory ocular disease in a human patient in need thereof. In another
embodiment, the present disclosure provides methods for the treatment of an ocular disease
in a veterinary patient in need thereof, including, but not limited to dogs, horses, cats,
rabbits, gerbils, hamsters, rodents, birds, aquatic mammals, cattle, pigs, camelids, and other
25 zoological animals.
In some embodiments of the compositions and methods disclosed herein, the
cyclosporine further comprises one or more additional active ingredients, e.g., active agents
selected from the group consisting of a resolvin or resolvin-like compound, a steroid (such
as a corticosteroid), and the like. In some embodiments, the additional active agent includes
30 a resolvin. In some embodiments, the additional active agent includes a corticosteroid. In
some embodiments, the additional active agent includes a resolvin and a corticosteroid. In
some embodiments, the additional active agent includes an antibiotic, for example one or
56
more antibiotics selected from the group consisting of azythromycin, ciprofloxacin,
ofloxacin, gatifloxacin, levofloxacin, moxifloxacin, besifloxacin, and levofloxacin. In some
embodiments, the additional active agent includes an antibiotic, for example one or more
antibiotics selected from the group consisting of azythromycin, ciprofloxacin, ofloxacin,
5 gatifloxacin, levofloxacin, moxifloxacin, besifloxacin, and levofloxacin; and a second of
such agents is a resolvin such as described herein (including without limitation compound
1001). In some embodiments, the active agent includes two or more active agents and one
of said active agents is an antiviral, for example one or more antivirals selected from the
group consisting of ganciclovir, trifluridine, acyclovir, famciclovir, valacyclovir,
10 penciclovir and cidofovir. In some embodiments, the active agent includes two or more
active agents and one of the active agents is an antibiotic, for example one or more antivirals
selected from the group consisting of ganciclovir, trifluridine, acyclovir, famciclovir,
valacyclovir, penciclovir and cidofovir; and a second of the active agents is a resolvin such
as described herein (including without limitation compound 1001).
15 Accordingly, in another aspect, provided is a method treating or preventing an ocular
disease or condition that includes locally administering a formulation of any of the aspects
or embodiments as disclosed herein. In some embodiments, the ocular disease is an anterior
segment disease. In some embodiments, the ocular disease is a posterior segment disease.
In some embodiments, the ocular disease is one or more selected from the group consisting
20 of dry eye syndrome, Sjogren's syndrome, uveitis, anterior uveitis (iritis), chorioretinitis,
posterior uveitis, conjunctivitis, allergic conjunctivitis, keratitis, keratoconjunctivitis, vernal
keratoconjunctivitis (VKC), atopic keratoconjunctivitis, systemic immune mediated
diseases such as cicatrizing conjunctivitis and other autoimmune disorders of the ocular
surface, blepharitis, scleritis, age-related macular degeneration (AMD), diabetic retinopathy
25 (DR), diabetic macular edema (DME), ocular neovascularization, age-related macular
degeneration (ARMD), proliferative vitreoretinopathy (PVR), cytomegalovirus (CMV)
retinitis, optic neuritis, retrobulbar neuritis, and macular pucker. In one embodiment, the
ocular disease is dry eye. In one embodiment, the ocular disease is allergic conjunctivitis.
In one embodiment, the ocular disease is age-related macular degeneration (AMD). In one
30 embodiment, the ocular disease is diabetic retinopathy.
The daily dose of the ophthalmic formulation, effective to reduce dry eye symptoms
and/or to improve tear film can be divided among one or several unit dose administrations.
A subject would use the product as needed, but generally, this would not be more than twice
57
a day and in many instances the product would be used only once a day. A preferred regimen
for the nanomicellar ophthalmic formulation of the present invention is one drop of O.09%
(w/w) solution per eye twice a day (approximately 12 hours apart).
EXAMPLES
5 To illustrate non-limiting embodiments of the present disclosure, the following
Examples were prepared.
EXAMPLE 1
Table 1: Cyclosporine nanomicellar ophthalmic formulation
Ingredients Amount
Cvclosporine 0.09wt %
hydrogenated 40 polyoxyl castor oil, 1.0wt%
octoxynol-40 0.05 wt%
sodium phosphate monobasic, 0.53 wt%
sodium phosphate dibasic, 0.47 wt%
sodium chloride, 0.05 wt%
povidone, 0.3 wt%
sodium hydroxide q.s. to adjust pH, ifrequired
hvdrochloric acid q.s. to adjust pH, ifrequired
water for injection q.s. to 100%
Cyclosporine nanomicellar ophthalmic solutions were prepared as follows. In
10 Example l(a), polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was melted at 55-
60°C with stirring at around 200 rpm. Cyclosporine A was added to the melted Kolliphor®
RH 40 at 55-60°C and the reaction mixture was mixed at the same temperature range until
complete dissolution. Following solubilization of the cyclosporine A, a surfactant
(octoxynol-40) was added under stirring and after 10 minutes of stirring, this non-aqueous
15 solution was delivered at 55-60°C to 90% water for injection. The temperature of the water
for injection was maintained at < 22°C. Sodium phosphate monobasic, sodium phosphate
dibasic, sodium chloride and povidone, were added to the bulk solution sequentially under
stirring until complete dissolution. Once all ingredients were completely solubilized in the
bulk solution, the volume was made up to 100% with water for injection to 1 L. In Example
20 l(b), the procedures of Example l(a) were followed, with the exception that the
cyclosporine A solution in Kolliphor® RH 40 at 55-60°C was added to water just after it
started showing turbidity.
58
The batches of both Examples l(a) and l(b) were filled into 3 piece 5 mL low density
polyethlylene ("LDPE") vials in an aseptic area. The vials were exposed to accelerated
temperatures of 40 °C and 30 °C in chambers (to accelerate particulate formation). The
samples were visually observed daily to see any sign of turbidity and/or visible particle
5 formation. The two batches were analyzed for critical quality parameters, such as, an assay
of cyclosporine, pH, osmolality, and micelle size, for which all was found well within
specification. The results are given in Table 2 below. Figure 1 shows photographs of the
stable batch (Example la) and the unstable batch (Example lb).
Table 2
Stability Condition Example 1 (a) Example 1 (b)
30°c Particle/turbidity
formation at 25 days
Particle/turbidity
formation at 11 days
Critical Quality Attributes
Assay of Cvclosporine 100.05 100.10
Total impurities 0.470 0.649
Ph 6.86 6.79
Osmolalitv 167 171
Particle Size
Zavg 16.29 16.95
PDI 0.148 0.172
The data in Table 2 illustrates that that when the batch is manufactured using a clear
CsA non-aqueous phase (cyclosporine in Kolliphor® RH 40 and octoxynol-40) and then
adding the mixture to a water phase, the batch remains stable for a longer time. However,
when the batch is manufactured using a turbid cyclosporine non-aqueous phase and then
adding the mixture to the water phase, the batch shows lower stability. This demonstrates
15 that a heterogeneous distribution of cyclosporine within micelles might facilitate nucleation
and particle formation in a finished product upon storage.
EXAMPLE2
The solution stability of cyclosporine A lots in polyoxyl 40 hydrogenated castor oil
20 (Kolliphor® RH 40) at 55-60°C is an important process parameter for a stable formulation.
It was also surprisingly found that the solubility behavior varied over the time of the storage,
as shown in Table 3.
59
Table 3: Solution stability of cyclosporine in Kolliphor® RH 40 at 55°C
Parameter
Solution stability of CsA in Kolliphor® RH 40 at 55°C
Initial data at
receipt of the
API
Current data
Study Study 1 Study 2 Study 3 Study 4 Study 5
Time to dissolve at
55°C (min) 20-30 45 40 165# 63 62
Total time for which the
solution remains clear
at 55°C (min) after API
is totally dissolved
(acceptance criteria is
NLT 75 min)
81*
60 90 7 99 102
52*
* Average of three studies results. # Some undissolved particles remained for a long time and
solution started to become slightly hazy very fast after complete solubility.
5 EXAMPLE3
Similarly, the solubility behavior study of cyclosporine A in polyoxyl 40
hydrogenated castor oil (Kolliphor® RH 40) was tested at 35°C. Table 4 shows the results
of a solubility behavior study of cyclosporine A in Kolliphor® RH 40 at 35°C. As can be
seen from Table 4, it was found that the solution stability at 35°C was comparatively higher
10 than the solution stability at 55°C.
Table 4: Comparative solution stability of cyclosporine A in amorphous form m
Kolliphor® RH 40 at 35°C and 55°C
Lot No. of CsA
3s0c ss0c
Solution stability
at 35°C SD Solution stability
at 55°C SD
Lot 7 239 10 92.17 14.77
Lot 8 304 12 99.33 4.62
Lot 9 248 17 79.67 9.29
Lot 10 125 0 49.67 17.79
Lot 11 62 0 23.25 1.50
Lot 12 235 7.64 74.33 4.93
Lot 17 135 0 46 0.00
Lot 18 114 0 44 0.00
Lot 19 80 0 47 0.00
60
As can be seen from Table 4, if cyclosporine A was not completely dissolved in
Kolliphor® RH 40 at 55°C, the temperature can be reduced to 35°C. Based on this data, it is
believed that the solution stability of cyclosporine A at 35°C was higher than that of 55°C.
The complete dissolution stage of cyclosporine A in Kolliphor® RH 40 was modified as
5 follows:
(i) API was dispersed at 55°C±2°C for 20 min and stirred for another 15 minutes;
(ii) the temperature was then decreased to 35°C±2°C from 55°C±2°C in less than 55
min (about 40 min), and the solution was stirred at 35°C±2°C until complete
dissolution of the API was achieved; and
(iii) octoxynol-40 was added with stirring and the non-aqueous phase was delivered
to water for injection at 35°C±2°C.
The changes in this dissolution process can accommodate lot to lot variability of the
solution stability of API in Kolliphor® RH-40 at 55 °C and also variability during the storage
as well.
15
EXAMPLE4
The solubility behavior of three cyclosporine forms as disclosed in Figure 2 were
studied in polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) at 55°C. The results are
shown in Table 5.
20 Table 5: Solubility of cyclosporine in Kolliphor® RH40 at 55 °C
Parameters
studied API type
CsA form with
characteristic
XRD peaks at 2-
theta (deg.) 6.9,
7.8, 9.4 and 15.9.
CsA form with
characteristic XRD
peaks at 2-theta
(deg.) 7.4, 8.7, 14.4
and 17.5
CsA form with
characteristic
XRD peaks at 2-
theta (deg.) 8.5,
9.3, 11.6 and
20.3.
Amorphous
API
Time taken to
dissolve at
55°C (min.)
15 14 min with haziness 15 min with
haziness 8
Time at which
haziness
appears (min.)
130 NA NA 92
61
Figures 3(a) to 3(d) are photographs of the results of the study reported in Table 5.
As can be seen from Table 5 above and from the associated figures, the CsA form
with characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7, 14.4 and 17.5 and the CsA form
with characteristic XRD peaks at 2-theta (deg.) 8.5, 9.3, 11.6 and 20.3 have much less
5 solubility as compared to the CsA form with characteristic XRD peaks at 2-theta (deg.) 6.9,
7.8, 9.4 and 15.9. During dissolution of cyclosporine in Kolliphor® RH 40 at 55°C, the CsA
form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9 may change into
less soluble forms, or amorphous cyclosporine may recrystallize into comparatively less
soluble forms. This conversion may be dependent on stresses to the system, including
10 temperature, long storage times at higher temperatures, and the like. To this end, the
cyclosporine ophthalmic formulation was exposed to higher temperatures and times and
PXRD data was taken from the precipitated part; it was found that the precipitate was close
to the CsA form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9, as
shown in Figure 4. Because this CsA form has lower solubility in water, its use may lead to
15 a seeding effect either at the initial stage or during storage at higher temperatures, thus
leading to batch failure.
In another attempt, the behavior of CsA in Kolliphor® RH 40 alone in absence of
water is determined. The CsA was dissolved in Kolliphor® RH 40 at 55°C and kept for a
longer time till precipitation occurred. The precipitated part was separated out and PXRD
20 was done. It was found that the cyclosporine has characteristic XRD peaks at 2-theta (deg.)
8.5, 9.3, 11.6 and 20.3, as shown in Figure 5. As a result, the process as disclosed in the
present disclosure, in which there is reduced exposure of cyclosporine to, for example,
Kolliphor® RH 40 at 55°C, can minimize this precipitation phenomenon which may be due
to a possible conversion of the cyclosporine to the less soluble CsA form. Thus, the present
25 disclosure provides for, among other things, improved storage stability.
EXAMPLES
Preparation method of the nanomicellar solution of Table 1
The polyoxyl 40 hydrogenated castor oil (Kolliphor® RH40) was heated to about 50-
30 60°C, until it liquefies, prior to introduction into the l0L glass vessel. The cyclosporine
(CsA) was added while maintaining the vessel temperature at 55±2°C and dissolved by
stirring at approximately 200-300 RPM for 75 minutes and visually inspected to ensure it is
62
a clear solution with no visible particles. The temperature was gradually lowered to 35°C.
Once it is completely dissolved, the temperature was increased to 55±2°C. The octoxynol-
40 is then added. If the octoxynol-40 solidified, it was heated at about 50-60°C until it
liquefies prior to its addition.
5 A portion (approximately 90%) of the Water for Injection (WFI) was charged into
the stainless-steel mixing tank and the temperature was maintained at 20-30°C throughout
the process. While stirring, the API mixture was added at 55±2°C to the mixing tank and
stirred for approximately 15 minutes while the remaining excipients were added in order of
sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, and
10 then polyvinylpyrrolidone.
After mixing for 15 minutes, the pH was checked and adjusted, if necessary, to 6.8±
0.2 using hydrochloric acid (IN) or sodium hydroxide (IN). The solution was adjusted to
the final volume with WFI and filtered through 0.2 μm filter.
15 EXAMPLE 6
Preparation method of the nanomicellar solution of Table 1
The polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was heated to about
50-60°C, until it liquefies, prior to introduction into the lOL glass vessel. The cyclosporine
(CsA) was added while maintaining the vessel temperature at 55±2°C for 20±2 minutes and
20 then stirred at approximately 200-300 RPM for 15 minutes. The temperature was reduced
gradually to 35°C under stirring and once it reached the temperature 35°C, it is stirred for
60±5 minutes. The octoxynol-40 was then added. If the octoxynol-40 solidified, it was
heated at about 50-60°C until it liquefies prior to its addition.
A portion (approximately 90%) of the Water for Injection (WFI) was charged into
25 the stainless-steel mixing tank and the temperature was maintained at 20-30°C throughout
the process. While stirring, the CsA mixture was added to the mixing tank at 35±2°C and
stirred for approximately 15 minutes while the remaining excipients were added in order of
sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, and
then polyvinylpyrrolidone.
63
After mixing for 15 minutes, the pH was checked and adjusted, if necessary, to 6.8±
0.2 using hydrochloric acid (IN) or sodium hydroxide (IN). The solution was adjusted to
the final volume with WFI and filtered through 0.2 μm filter.
5 EXAMPLE7
Preparation method of the nanomicellar solution of Table 1 - process under high speed
The polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was heated to about
50-60°C, until it liquefies, prior to introduction into the lOL glass vessel. The cyclosporine
(CsA) was added while maintaining the vessel temperature at 55±2°C for 20±2 minutes and
10 stirred at approximately >450 RPM for 15 minutes. The temperature was reduced gradually
to 35°C under stirring and stirred for 60±5 minutes. The octoxynol-40 was then added. If
the octoxynol-40 solidified, it was heated at about 50-60°C until it liquefies prior to its
addition.
A portion (approximately 90%) of the Water for Injection (WFI) was charged into
15 the stainless-steel mixing tank and the temperature was maintained at 20-30°C throughout
the process. While stirring, the CsA mixture was added to the mixing tank at 35±2°C and
stirred for approximately 15 minutes while the remaining excipients were added in order of
sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, and
then polyvinylpyrrolidone.
20 After mixing for 15 minutes, the pH was checked and adjusted, if necessary, to 6.8±
0.2 using hydrochloric acid (IN) or sodium hydroxide (IN). The solution was adjusted to
the final volume with WFI and filtered through 0.2 μm filter.
EXAMPLES
25 Preparation method of the nanomicellar solution of Table 1 - process at higher temperature
The polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was heated to about
50-60°C, until it liquefies, prior to introduction into the 10L glass vessel. The temperature
was increased to 127-130°C. The cyclosporine (CsA) was added while maintaining the
vessel temperature at 127-130°C and stirred at approximately 200-300 RPM for complete
30 dissolution. The octoxynol-40 was then added. If the octoxynol-40 solidified, it was heated
at about 50-60°C until it liquefies prior to its addition.
64
A portion (approximately 90%) of the Water for Injection (WFI) was charged into
the stainless-steel mixing tank and the temperature was maintained at 20-30°C throughout
the process. While stirring, the CsA mixture was added at 127-130°C to the mixing tank and
stirred for approximately 15 minutes while the remaining excipients were added in order of
5 sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, and
then polyvinylpyrrolidone.
After mixing for 15 minutes, the pH was checked and adjusted, if necessary, to 6.8±
0.2 using hydrochloric acid (IN) or sodium hydroxide (IN). The solution was adjusted to
the final volume with WFI and filtered through 0.2 μm filter.
EXAMPLE9
The polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was heated to about
50-60°C, until it liquefies, prior to introduction into the 10L glass vessel. The temperature
was increased to 127-130°C. The octoxynol-40 was then added. If the octoxynol-40
15 solidified, it was heated at about 50-60°C until it liquefies prior to its addition. The mixture
was stirred for 20 minutes at 127-130°C. The cyclosporine (CsA) was added while
maintaining the vessel temperature at 127-130°C and stirred at approximately 200-300 RPM
using a stirrer for complete dissolution.
A portion (approximately 90%) of the Water for Injection (WFI) was charged into
20 the stainless-steel mixing tank and the temperature was maintained at 20-30°C throughout
the process. While stirring, the CsA mixture was added at 127-130°C to the mixing tank and
stirred for approximately 15 minutes using both an overhead and bottom stirrer while the
remaining excipients are added in order of sodium phosphate monobasic, then sodium
phosphate dibasic, then sodium chloride, and then polyvinylpyrrolidone.
25 After mixing for 15 minutes, the pH was checked and adjusted, if necessary, to 6.8±
0.2 using hydrochloric acid (IN) or sodium hydroxide (IN). The solution was adjusted to
the final volume with WFI and filtered through 0.2 μm filter.
EXAMPLE 10
30 Stability Studies
65
The nanomicellar ophthalmic formulation of Example 6 was tested after being stored
at 25°C/40%RH for 6 months.
The formulation was tested for change in appearance, pH, osmolality, viscosity,
Cyclosporine assay by HPLC method II, micelle size determination by Laser light scattering
5 method and particulate matter presence.
Table 6:
Tests Specifications Initial 3M 6M
Appearance Clear colorless solution,
essentially free from visible
particulate matter
Complies Complies Complies
pH 6.5-7.2 6.8 6.8 6.8
Osmolalitv l 60- l 90mOsmol/Kg 169 166 175
Viscosity 1.3 -2.0 cP 1.4 1.4 1.4
HPLC method II
Cyclosporine assay 95.0 -105.0% oflabel claim 97.8 99.2 98.2
Total impurities g_0¾area 0.59 0.76 0.83
Micelle size
determination
Zavg: 13-16nm 15 14 14
Pdl :S0.2 0.13 0.15 0.10
Particulate matter
(USP 788)
The average number of
particles present in the units
does not exceed:
Stage 1 : by method 1 - Light
obscuration
6000 per contained 2::l0μm NA NA NA
600 per container 2::25μm NA NA NA
Stage 2: method 2- Membrane
. .
m1croscop1c
6000 per contained 2::l0μm NA NA NA
600 per container 2::25μm NA NA NA
Sterility No evidence of microbial
growth
Sterile NA NA
As observed from the Table 6 data, the formulation was found to be both chemically
and physically stable.
66
EXAMPLE 11
Stability Studies
The nanomicellar ophthalmic formulation of Example 8 was tested after being stored
at 25°C/40%RH for 24 months, at 30°C /35%RH for 6 months and at 40°C /25%RH for 6
5 months.
Table 7:
rrests Specification 25°C/40%RH
OM lM 3M 6M 12M 24M
k\ppearance
Clear,
colorless
solution,
essentially free
from visible
particulate
matter.
Complies Complies Complies Complies Complies Complies
k\ssay of
Cvclosporine
90.0-110% of
LC 93.32 93.27 94.69 94.32 93.48 93.98
Pnkimp@
RT 0.36 NMT0.7% 0.145 0.19 0.135 0.221 0.23 0.217
RS Known Imp.
Pnkimp@
RT 0.55 NMT0.7% 0.057 0.195 0.048 0.101 0.206 0.233
Pihydro CsA +CsA
K,7 NMT0.7% 0.265 0.231 0.260 0.283 0.281 0.249
CsAD NMT0.7% 0.082 0.07 0.071 0.037 0.042 0.08
OCso-CsA A NMT0.7% 0.516 0.123 0.180 0.149 0.154 0.207
CsAH NMT0.7% ND ND ND ND ND QL(<0.05%)
aighest UnKnown NMT0.1% 0.05
RRT: 1.32 0.099 0.080
2.07RRT 0.081 0.074 0.092
lfotal Impurities NMT2% 1.481 1.471 0.969 1.398 1.452 1.135
hrticle size
tAVGnm Bet 13-16 nm 16.08 14.03 13.7 14.38 13.66 14.38
rm. NMT0.2 0.146 0.115 0 0.140 0.095 0.150
pH 6.5-7.2 6.92 b.87 6.74 6.82 6.83 6.83
Psmolality 160-190
mOsmol/kg 163 NA 168 168 6.83 172
hrticulate
Matter
1<789>
l0μm NMT 50/ml NA NA 3.3 39.0 73.7 9.1
25μm NMT 5/ml NA NA 0.0 1.3 3.3 0.3
50μm NMT2/ml NA NA 0.0 0.0 0.0 0.1
67
Table 8:
rrests Specification 30°C /35%RH 40°C/25%RH
lM 3M 6M lM 3M 6M
k\ppearance
Clear,
colorless
solution,
essentially free
from visible
particulate
matter.
Complies Complies Complies Complies Complies Complies
k\ssay of
Cyclosporine
90.0-110% of
LC 93.16 94.62 92.6 93.09 94.36 91.18
RS Known Imp.
Unkimp@
RRT 0.36 NMT0.7% 0.183 0.149 0.226 0.239 0.215 0.367
Unkimp@
RRT 0.55 NMT0.7% 0.152 0.047 0.12 0.257 0.048 0.217
Dihydro CsA
+CsAV NMT0.7% 0.229 0.257 0.298 0.225 0.289 0.354
CsAD NMT0.7% 0.061 0.063 0.063 0.073 0.054 0.046
Iso-CsA A NMT0.7% 0.12 0.195 0.196 0.138 0.215 0.186
CsAH NMT0.7% ND ND ND ND ND ND
Highest
UnKnown NMT0.1% 0.077 0.070
RRT0.48 0.098
0.085
RRT:2.08
0.105
0.48RRT 0.127
Total Impurities NMT2% 1.376 0.988 1.578 1.629 1.175 2.076
hrticle Size
AVG nm Bet 13-16
nm 14.09 13.9 17.28 13.95 13.7 14.12
rm. NMT0.2 0.112 0.10 0.245 0.103 0.093 0.125
i:,H 6.5-7.2 6.87 6.74 6.82 6.86 6.73 6.8
bsmolality 160-190
mOsmol/kg NA 168 168 NA 169 169
hrticulatp
Matter
1<789>
lOμm NMT
50/ml NA 2.7 1.7 NA 2.3 39.0
25μm NMT 5/ml NA 0.0 0.0 NA 0.0 1.3
50μm NMT2/ml NA 0 0.0 NA 0.0 0.0
Unless indicated otherwise, the documents mentioned herein are incorporated by
reference in their entirety.
68
Even though certain specific embodiments are thoroughly described in the present
application, it should be understood that the same concepts disclosed with respect to those
specific embodiments are also applicable to other embodiments. Furthermore, individual
elements of the formulations and methods disclosed herein are described with reference to
5 particular embodiments only for the sake of convenience. It should be understood that
individual elements of the formulations and methods disclosed herein are applicable to
embodiments other than the specific embodiments in which they are described.
In addition, it should be understood that the scope of the present disclosure is not
limited to the above-described embodiments, and those skilled in the art will appreciate that
10 various modifications and alterations are possible without departing from the scope of the
present disclosure. For example, the batch sizes may be altered by a person having ordinary
skill in the art while staying within the present disclosure.
69
WE CLAIM:
1. A stable nanomicellar ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine, adding octoxynol-40; and
c) mixing the resulting mixture with the aqueous vehicle.
2. A stable nanomicellar ophthalmic formulation of claim 1 comprising a cyclosporine
form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4 and 15.9.
3. A stable nanomicellar ophthalmic formulation of claim 1 comprising an amorphous
form of cyclosporine.
4. A method of making a stable nanomicellar ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the method comprises the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine, adding octoxynol-40; and
c) mixing the resulting mixture with the aqueous vehicle.
5. A method of making a stable nanomicellar ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the method comprises the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) keeping mixture A under vacuum to remove foam;
70
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine;
d) adding octoxynol-40; and
e) mixing the resulting mixture with the aqueous vehicle.
6. A method of making a stable nanomicellar ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the method comprises the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) keeping mixture A under vacuum to remove foam;
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine;
d) adding octoxynol-40; and
e) mixing the resulting mixture with the aqueous vehicle.
7. A stable nanomicellar ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of
127-130°C.
8. A stable nanomicellar ophthalmic formulation of claim 7, wherein the formulation
comprises a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9.
9. A stable nanomicellar ophthalmic formulation of claim 7, wherein the formulation
comprises an amorphous form of cyclosporine.
71
10. A stable nanomicellar ophthalmic formulation of claim 7, wherein the formulation
comprises cyclosporine in a form having characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7,
14.4 and 17.5.
11. A stable nanomicellar ophthalmic formulation of claim 7, wherein the formulation
comprises cyclosporine in a form with characteristic XRD peaks at 2-theta (deg.) 8.5, 9.3,
11.6 and 20.3.
12. A stable nanomicellar ophthalmic formulation comprising:
cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous
vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a
temperature of 127-130°C to form a mixture A;
b) adding cyclosporine to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of
127-130°C.
13. A stable nanomicellar ophthalmic formulation of claim 7, wherein the formulation
comprises a cyclosporine form with characteristic XRD peaks at 2-theta (deg.) 6.9, 7.8, 9.4
and 15.9.
14. A stable nanomicellar ophthalmic formulation of claim 7, wherein the formulation
comprises an amorphous form of cyclosporine.
15. A stable nanomicellar ophthalmic formulation of claim 7, wherein the formulation
comprises cyclosporine in a form having characteristic XRD peaks at 2-theta (deg.) 7.4, 8.7,
14.4 and 17.5.
16. A stable nanomicellar ophthalmic formulation of claim 7, wherein the formulation
comprises cyclosporine in a form with characteristic XRD peaks at 2-theta (deg.) 8.5, 9.3,
11.6 and 20.3.
17. A stable nanomicellar ophthalmic formulation comprising:
0.09 wt % cyclosporine,
about 1.0 wt % hydrogenated 40 polyoxyl castor oil,
72
about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) keeping mixture A under vacuum to remove foam;
c) optionally, lowering the temperature of mixture A to a temperature of
35°C±2°C prior to the complete dissolution of the cyclosporine;
d) adding octoxynol-40; and
e) mixing the resulting mixture with the aqueous vehicle.
18. A stable nanomicellar ophthalmic formulation, prepared by a method comprising the
steps of:
a) m1xmg cyclosporine with hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine, adding octoxynol-40; and
c) then mixing the resulting mixture with an aqueous vehicle at 35°C±2°C,
wherein the mixture A is lowered to a temperature of 35°C±2°C in less than 65
minutes.
19. A stable nanomicellar ophthalmic formulation of claim 18 wherein the mixture A is
lowered to a temperature of 35°C±2°C in 40-50 minutes.
20. A stable nanomicellar ophthalmic formulation of claim 18 wherein the temperature
of mixture A is lowered to 35°C±2°C and then stirring mixture A for 60-70 minutes at a
temperature of 35°C±2°C.
21. A stable nanomicellar ophthalmic formulation comprising:
0.09 wt % cyclosporine,
about 1.0 wt % hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
73
an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 127-130°C until completely dissolved to form a mixture A;
b) adding octoxynol-40 to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
22. A stable nanomicellar ophthalmic formulation of claim 21, further comprising:
about 0.20-0.550 wt% sodium phosphate monobasic,
about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
about 0.3 wt% povidone, and
sodium hydroxide/hydrochloric acid to adjust the pH
23. A stable nanomicellar ophthalmic formulation comprising:
0.09 wt % cyclosporine,
about 1.0 wt % hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40, and
an aqueous vehicle,
wherein the ophthalmic formulation is made by a method comprising the steps of:
a) mixing the hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a
temperature of 127-130°C to form a mixture A;
b) adding cyclosporine to mixture A at 127-130°C; and
c) mixing the resulting mixture with the aqueous vehicle at a temperature of at
127-130°C.
24. A stable nanomicellar ophthalmic formulation of claim 23, further comprising:
about 0.20-0.550 wt% sodium phosphate monobasic,
74
about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
about 0.3 wt% povidone, and
sodium hydroxide/hydrochloric acid to adjust the pH
25. A method of making a stable nanomicellar ophthalmic formulation comprising:
0.09 wt % cyclosporine,
about 1.0 wt % hydrogenated 40 polyoxyl castor oil,
about 0.05 wt% octoxynol-40,
about 0.20-0.550 wt% sodium phosphate monobasic,
about 0.23-0.465 wt% sodium phosphate dibasic,
about 0.05 wt% sodium chloride,
about 0.3 wt% povidone,
sodium hydroxide/hydrochloric acid to adjust the pH, and
water for injection,
wherein the method comprises the steps of:
a) mixing the cyclosporine with the hydrogenated 40 polyoxyl castor oil at a
temperature of 55°C±2°C or above to form a mixture A;
b) lowering the temperature of mixture A to a temperature of 35°C±2°C prior
to the complete dissolution of the cyclosporine, adding octoxynol-40 to form an API
mixture;
c) adding the API mixture to Water for Injection (WFI);
d) adding remaining excipients to step (c) in the order of sodium phosphate
monobasic, then sodium phosphate dibasic, then sodium chloride, and then
polyvinylpyrrolidone; and,
e) adjusting the pH to 6.5 to 7.2 and bringing to the final volume with WFI.
75
26. The method of claim 22, wherein the mixing cyclosporine in step (a) occurs at
200- 300 RPM
27. The method of claim 1, wherein the temperature of mixture A to 35°C±2°C in 40-
50 minutes.
28. The method of claim 1, wherein mixture A 1s stirred for 60±5 minutes at a
temperature of 35°C±2°C.
29. A stable nanomicellar ophthalmic formulation prepared by the method as described
in any of the preceeding claims.
30. The stable nanomicellar ophthalmic formulation of claim 25, wherein the osmolality
of the formulation is between about 150 to about 200 mOsmol/kg.
31. The stable nanomicellar ophthalmic formulation of claim 25, wherein the
formulation comprises a cyclosporine form with characteristic XRD peaks at 2-theta (deg.)
6.9, 7.8, 9.4 and 15.9.
32. The stable nanomicellar ophthalmic formulation of claim 25, wherein the
formulation comprises an amorphous form of cyclosporine.
33. The stable nanomicellar ophthalmic formulation of claim 25, wherein the
formulation is substantially free of a cyclosporine form with characteristic XRD peaks at 2-
theta (deg.) 7.4, 8.7, 14.4 and 17.5.
34. The stable nanomicellar ophthalmic formulation of claim 25, wherein the
formulation is substantially free of a cyclosporine form with characteristic XRD peaks at 2-
theta (deg.) 8.5, 9.3, 11.6 and 20.3