The present invention generally relates to a vitamin 03 (cholecalciferol) composition and method
of preparation thereof. Preferably the present invention discloses a nano-emulsion formulation of
vitamin 03 wherein the dosage comprising high dose of the vitamin 03 with improved drug
absorption rate and bio-availabi I ity.
BACKGROUND OF THE INVENTION:
Vitamin D (cholecalciferol) is a fat-soluble vitamin playing a vital role in human physiology as a
positive regulator of calcium homeostasis. Vitamin 0 deficiency is prevalent worldwide. This
deficiency has many consequences which are still being explored, apart from the well-known
skeletal complications. Vitamin 0 insufficiency is not only linked to bone disease but also to
several non-skeletal conditions, including type 2 diabetes mellitus, cardiovascular disease, chronic
lung di~ease, tuberculosis and upper respiratory infections. Our Indian diet generally fails to satisfy
the daily requirement of Vitamin 0 for a normal adult.
About 90% of the required Vitamin 0 is synthesized in the skin under sun exposure The prevalence
of Vitamin D deficiency is reported worldwide, both in sunshine deficient and sunshine sufficient
countries. Still, it is the most underdiagnosed and undertreated nutritional deficiency in the world.
In India, more than 90% of apparently healthy Indians have subnormal 25(0H) D levels. To
maintain sufficient vitamin D level, apart from sunlight and food containing vitamin 0,
supplementation with vitamin Dis also required. Very few foods naturally contain vitamin D, and
foods that are fortified with vitamin Dare often inadequate to satisfy either a child's or an adult's
vitamin 0 requirement.
Vitamin D is a group of secosteroids with a 4-ringed cholesterol backbone, has two major forms:
(a) Vitamin 03, i.e., cholecalciferol and (b) Vitamin 02, i.e., ergocalciferol. Vitamin D, in general,
refers to Vitamin 03. Calciferol is a collective term refers to both forms of Vitamin D. In India
current recommendations for correction of vitamin D level, is by giving 60,000 IU of oral Vitamin
0 on a weekly basis for 8 weeks.
Correction of vitamin 0 insufficiency is commonly achieved using oral vitamin D supplements .
The Endocrine Society guidelines suggest that daily intake of I ,500 to 2,000 international units
Page 2 of 16
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(IU) of vitamin Dis necessary to achieve serum 25(0H)D concentrations consistently >30 ng/mL
in adults. However, adherence to daily doses has been reported to be low in several large clinical
trials. Poor adherence has been associated with difficulty swallowing combined vitamin D/calcium
tablets, gastrointestinal (G I) side-effects, the number of concurrent treatments a patient is
receiving, and the patient's attitude towards vitamin D supplementation.
Currently available oral Vitamin D supplements comprise either vitamin 02 (ergocalciferol) or
vitamin 03. As per the market scenario, only two preparations contain vitamin 02 (ergocalciferol).
More than 99.9% of the available preparations contain vitamin 03 in the form of alfacalcidiol (25
hydroxycholecalciferol), calcitriol (I ,25 dihydroxycholecalciferol) or cholecalciferol (inactive
vitamin D). Most of the preparations contain calcitriol · (46.5%) or alfacacidiol (43.02%).
Approximately I 0% of preparations contain cholecalciferol (I 0.5%).
Table I: Currently marketed dosage forms of cholecalciferol
Product Manufacturer Form of the Dosage form Amount/dosag
Name Vitamin D e form
Rocaitrol Abott Healthcare Pvt. Cholecalciferol Capsules 0.25 Jlg
Ltd.
Vitanova SG Zuventus Healthcare Ltd. Cholecalciferol Capsules 60000 IU
Quente 03 Stanley Healthcare Cholecalciferol Capsules 60000 IU
Vital cal Genesis Biotec Inc Cholecalciferol Tablets 60000 IU
Hira 03 Hirai Lab Ltd. Cholecalciferol Tablets 60000 IU
Arachitol Solvay Pharma India Pvt. Cholecalciferol Injection 600000 IU
Ltd.
Fovit 03 Forgo Pharmaceuticals Cholecalciferol Injection 600000 IU
Pvt. Ltd.
03 UP Abott Healthcare Pvt. Cholecalciferol Granules 60000 IU
Ltd.
Caldikind. Mankind Pharmaceutical Cholecalciferol Granules 60000 IU
Pvt. Ltd.
Calotec 03 Lupin Laboratories Cholecalciferol Powder 60000 IU
Page 3 of 16
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It can be seen from above Table I that solid dosage form ofthe cholecalciferol contains 60000 IU
of Vitamin D2 and are formulated for quick or immediate release in the gastrointestinal tract. No
oral dosage forms of the Cholecalciferol comprises higher amount of cholecalciferol than 60000
IU.
Indian patent application IN934/DEL/2015 claims liquid palatable dosage forms of Vitamin D
comprising hydrogenated castor oil as a solubilizer and polysorbate 80 as a surfactant. Each 5 ml
of the liquid dosage form contains 120000 IU of vitamin D.
Patent application W02008134512 claimed controlled release formulation ofVitamin D. As per
the patent specification each soft gelatin capsule contain 10000 IU of Vitamin D
In order to solve the above problems, the present invention is intended to provide a nano-emuision
formulation of vitamin 03 wherein the dosage comprising high dose of the vitamin D3 with
improved drug absorption rate and bio-availability along with reduced dosing frequency.
Therefore if Vitamin 03 is given as a single, large oral dose will demonstrate higher adherence
rates compared with daily or weekly dosing regimens, and may have the potential to yield sustained
improvements in serum 25(0H)D.
No prior ~rt reported in the literature linked with nano-emulsion formulation of vitamin D3
comprising higher dose/amount of vitamin D3. The present invention related to nano-emuision
formulation of vitamin 03 which comprises not less than 240000 IU of cholecalciferol per capsule
or liquid dosage form containing not less than 240000 IU of cholecalciferol per 5 ml, which may
reduce the dosing frequency of vitamin D3 to monthly or yearly from daily or weekly with
improved drug absorption rate and bio-availability.
SUMMARY OF THE INVENTION:
The present invention generally relates to a vitamin D3 (cholecalciferol) composition and method
of preparation thereof.
Preferably the present invention relates to a novel pharmaceutical dosage form of the vitamin D3
and method of preparation thereof.
Page 4 of 16
"
)
More preferably the present invention relates to a pharmaceutical dosage form ofthe vitamin 03
and method of preparation thereof.
More specifically the invention discloses a nano-emulsion formulation of vitamin 03 wherein
vitamin 03 is in the form of nano-oil globules dispersed in aqueous phase.
The present invention also relates to a unit dosage form of the vitamin 03 comprising higher dose
of the vitamin 03, wherein the amount of the Vitamin 03 contained in the formulation is not less
·than. 240000 IU of cholecalciferol per capsule or liquid dosage form containing not less than.
240000' IU of cholecalciferol per 5 ml,
The present invention also relates to a vitamin 03 formulation .possessing higher bio-availability
with improved drug absorption rate than marketed preparations.
BRIEF DESCRIPTION OF FIGURES:
Figure 1: In-vitro Drug Release Profile
Figure 2: Comparison of in vivo Pharmacokinetic profile of Vitamin 03 PBL formulation (Liquid
oral) and Marketed Formulation (Liquid oral formulation)
Figure 3: Comparison of in vivo Pharmacokin' etic profile of Vitamin 03 PBL formulation (solid·
Oral) and Marketed Formulation (Solid oral and Injectable formulation)
DETAILED DESCRIPTION OF THE INVENTION:
Hereinafter, a preferred embodiment of the present invention will be described. It should be noted
that those of ordinary skill in the art who, without departing from the principles of the present
invention may also be made to a number of improvements and retouch, these improvements and
modifications should also be regarded as the protection scope ofthe present invention
The present invention is directed to pharmaceutical compositions compnsmg vitamin 0
compounds in emulsion formulations.
Preferred form of the vitamin 0 included in the formulation ofthe invention is chlecalciferol.
Page 5 of 16
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However Examples of vitamin 0 compounds suitable for formulations of the present invention
include, without limitation are ergocalciferol, alfacalcidiol (25 hydroxycholecalciferol), calcitriol
(I ,25 dihydroxycholecalciferol), la,2- dihydroxyvitarnin 04, la,24-dihydroxy vitamin 02, Ia
hydroxyvitamin 03 (a-calcidol), la,25-dihydroxyvitamin 02, la,25-dihydroxyvitamin 04, 1,24,25-
dihydroxyvitamin 02, seocalcitol (EB-1 089), calcipotriol, 22-oxacalcitriol (maxacalcitol),
fluorinated compounds such as falecalcitriol, and 19-nor compounds such as paricalcitol.
The amount of selected therapeutic is not critical to the present invention and may be varied to
achieve the desired therapeutic response for a particular patient. The amount of active therapeutic
agent in the formulations ofthe invention will be dependent, in part, on the solubility ofthe specific
surfactant used and its intended use. Those skilled in the arts can adjust the ratios without undue
experimentation. As one of the objectives ofthe present invention is to reduce the dosing frequency
via increasing the drug content per unit dosage form, present invention comprises a soft gelatin
capsule of Vitamin D containing not less than 240000 IU ( <2% w/w) of cholecalciferol per capsule
or liquid dosage form containing not less than 240000 IU ( <0.1% w/v) of cholecalciferol per 5 mi.
According to one aspect of the invention, pharmaceutical compositions are provided comprising a
lipophilic phase component, hydrophilic phase, one or more surfactants, and an active vitamin 0
compound; wherein said composition is a nano-emulsion, which is to be filled in soft gelatin
capsule or administered as a liquid dosage form.
The pharmaceutical compositions of the present invention comprise a lipophilic phase component.
Suitable components for use as lipophilic phase components include any pharmaceutically
acceptable solvent which is non- miscible with water, The lipophilic phase component may
comprise mono-, di- or triglycerides. Mono-, di- and triglycerides that may be used within the
scope ofthe invention include those that are derived from C6, Cs, C10, C12, C14, C16, C1s, C2o and
C22 fatty acids. Exemplary diglycerides include, in particular, diolein, dipalmitolein, and mixed
caprylin-caprin diglycerides. Preferred triglycerides include vegetable oils, fish oils, animal fats,
hydrogenated vegetable oils, partially hydrogenated vegetable oils, synthetic triglycerides,
modified triglycerides, fractionated triglycerides, medium and long-chain triglycerides, structured
triglycerides, and mixtures thereof. Among the above-listed triglycerides, preferred triglycerides
include almond oil; babassu oil; borage oil; blackcurrant seed oil; canola oil; castor oil coconut
oil; corn oil; cottonseed oil; evening primrose oil; grapeseed oil groundnut oil; mustard seed oil;
Page 6 of 16
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olive oil; palm oil; palm kernel oil; peanut oil rapeseed oil; safflower oil; sesame oil; shark liver
oil; soybean oil; sunflower oil hydrogenated castor oil; hydrogenated coconut oil; hydrogenated
palm oil hydrogenated soybean oil; hydrogenated vegetable oil; hydrogenated cottonseed and
castor oil; partially hydrogenated soybean oil; partially soy and cottonseed oil; glyceryl tricaproate;
glyceryl tricaprylate; glyceryl tricaprate; glyceryl triundecanoate; glyceryl trilaurate; glyceryl
trioleate; glyceryl trilinoleate; glyceryl trilinolenate; glyceryl tricaprylate/caprate; glyceryl
tricaprylate/caprate/laurate; glyceryl tricaprylate/caprate/linoleate; and glyceryl
tricaprylate/caprate/stearate.
A preferred triglyceride is the medium chain triglyceride. More preferred medium chain
triglycerides are caprylic-capric acid triglycerides. caprylic-capric acid triglycerides are known
and commercially available under the trade name MYRITOL, including the product MYRITOL
813 Especially preferred as lipophilic phase component in the invention. Amounts of such
triglyceride present in the formulations of the present invention include from about I 0% to about
20% w/w.
Suitable surfactants for the formulations of the present invention include non ionic surfactants. A
non-ionic surfactant is one where the hydrophilic part of the surfactant carries no charge but
derives its water solubility from highly polar groups such as hydroxyl or polyoxyethylene groups.
Nonionic surfactants generally include, but are not limited to, the polyoxyalkylenes dextrans, fatty
acid esters of saccharose, fatty alcohol ethers of oligoglucosides (e.g., the akylpolyglucosides such
as TRITON™), fatty acid esters of glycerol (e.g., glycerol mono/distearate or glycerol
monolaurate), and polyoxyethylene type compounds (e.g., POE, PEG, PEG, SOLUTOL™
CREOMOPHOR™s, MACROGOL, CARBO WAX, POL YOXYL). The later also include
polyethoxylated fatty acid esters of sorbitan (e.g., polysorbates, such as TWEEN™s, SPAN™s),
fatty acid esters of (polyethylene oxide) (e.g., polyoxyethylene stearates), fatty alcohol ethers of
poly(ethylene oxide) (e.g., polyoxyethylated lauryl ether), alkylphenol ethers of poty(ethylene
oxide) (e.g.,polyethoxylated octylphenol), polyoxy~thylene-polyoxypropylene block copolymers
(also known as poloxamers, such as "Piuronic"), and ethoxylated fats and oils (e.g., ethoxylated
castor oil, or polyoxyethylated castor oil, also known as polyethylene glycolglyceryl
triricinoleate). Mixtures ofsurfactants are also within the scope ofthe invention. Amounts of such
Page 7 of 16
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surfactant present in the formulations of the present invention include from about 40% to about
70% w/w.
Pharmaceutical compositions of the present invention may further comprise a hydrophilic phase
component. The hydrophilic phase component may comprise, e.g., a pharmaceutically acceptable
C 1-s alkyl or tetrahydrofurfuryl di- or partial-ether of a low molecular weight mono- or poly-oxyalkanediol.
In a preferred embodiment, the hydrophilic phase component comprises I ,2-propylene
glycol. Amounts of such hydrophilic phase component present in the formulations of the present
invention include from about I 0% to about 20% w/w.
The pharmaceutical compositions of the present invention may further comprise one or mor.e
additives. Additives that are well known in the art include, e.g., detackifiers, anti-foaming agents,
buffering agents, antioxidants (e.g., ascorbyl palmitate, butyl hydroxy anisole (BHA), butyl
hydroxy toluene (BHT) and tocopherols, e.g., a-tocopherol (vitamin E)) , preservatives, chelating
agents, viscomodulators, tonicifiers, flavorants, colorants odorants, opacifiers, suspending agents,
binders, fillers, plasticizers, lubricants, and mixtures thereof. The amounts of such additives can
be readily determined by one skilled in the art, according to the particular properties desired .
•
In order to obtain good stability, high efficiency and high bio-availability long-acting compound
vitamin 03 nano-emulsion to the body system, the present invention and an amount of each
component were carefully screened, and the order of addition and manner of addition of each
component is adjusted.
An object of the present invention provides a vitamin' 03 nano-emulsions, the product is clear,
thermodynamically good stability; has sustained release, long lasting drug efficacy; nanoemulsion
particles large specific surface area, can obviously improve the drug absorption rate and
bioavailability; system compatibility is good, in addition to the vitamins, reproducibly formulated
in a variety of beneficial ingredients. The product is primarily used to supplement the body needs
ofvitamins, prevention and treatment of vitamin deficiencies
Present invention discloses o/w type of nano-emulsion of the vitamin 0 wherein vitamin 0 is
present in the form of the nano-globules in dispersed in aqueous external phase. Size reduction or
Page 8 of 16
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50 percent { d50) of vitamin 0 globules < I oo· nm is accountable for its enhanced bioavailability
in the present invention.
Compound vitamin D3 nano-emulsion ofthe present invention is stablewith an average diameter
of less than I 00 nm; at 25 oc. The present vitamin 03 nano-emulsion particle size is small, in the
range of I 00 nm or less, have a powerful molecular surface effect, vitamin molecule stable, not
subject to oxidation damage, stable thermodynamic properties. Nano-sized vitamin greatly
improves the solubility ofthe vitamin in water, dispersible and dialysis performance, changes the
vitamins in vivo physiological and biochemical processes, and eliminates the inter-individual
differences in the absorbed and utilized, may be up to 100% absorption, bioavailability reaches
90% or more.
The present invention of vitamin 03 nano-emulsion with water-miscible and emulsion droplets
will not precipitate at an arbitrary ratio, vitamin level can be reduced, reducing the overall cost to
25%, cost-effective, in actual use shown great advantages.
The present invention also discloses a method of preparing solid dosage or liquid dosage form of
vitamin 03 where the vitamin 03 is present as nano-sized droplets in the dispersant.
The embodiments of the present invention are further illustrated by the following examples which
are provided merely to be exemplary of the invention and do not limit the scope of the invention.
Certain modifications and equivalents will be apparent to those skilled in the art and are intended
to be included within the scope of the invention .
• Example 1: Composition ofVitamin 03 Soft Gelatin Capsules and its Process of preparation:
Preparation ofNano-emulsion of Vitamin D
Table 1: Composition
S. No. Ingredients
Quantity Per
Capsule (Mg)
1. Cholecalciferol 6.0
2 . Miglyol Oil 812 21.43
3. Cremophor RH40 I 00.525
Page 9 of 16
4. Propylene Glycol 21.43
5. Tocopherol 0.015
1. Manufacturing Procedure
1.1 Miglyol Oil812 and propylene glycol were taken in a vessel and stat1ed purging Nitrogen gas
(99.90% pure). Cholecalciferol was dissolved in it.
1.2 Added and dissolved Cremophor RH40 and tocopherol in vessel of step I and mixed with
continuous stirring in an area at a temperature below 25°C, protected from light and moisture.
Increased the speed of the stirrer, so that a vortex is formed and continued stirring for about I
hour.
1.3 Filtered the medicament solution of step 1.2 through nylon filter cloth of mesh size 160.
1.4 Stored the filtrate in closed S.S. container after nitrogen purging at a temperature below 25°C,
protected from light and moisture.
Above formulated nano-emulsion of Example I was filled in soft gelatin capsule shells.
Example 2
Each soft gelatin capsule contains Cholecalciferol (Vitamin 03) ................ 240000 IU
Table 2: CAPSULE SHELL COMPOSITION
S. NO. INGREDIENTS
PERCENT
COMPOSITION
I. Gelatin 160 bloom strength geletia 42.38
2. Gelatin (low mol wt) Byco A 4.24
3. Glycerol 17.15
4. colour quinoline yellow supra 0.05
5. Water 36.18
Page 10 of 16
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2. Preparation of gelatin mass
2.1 Charged purified water and glycerin in to the gelatin melting reactor with the help of
Vacuum. Heated the reactor to 70-75°C.
2.2 Added gelatin to the melting reactor with the help of vacuum. Continued stirring with the help
of stirrer at 30-50 RPM for 60 minutes, at a temperature of 70-75°C till homogenous paste
was formed. Applied 600-700 mm Hg of vaccum with continuous stirring for 30-60 minutes,
till gelatin mass was free from air bubble.
2.3 Discharged the gelatin mass from gelatin reactor into gelatin mass storage vessel through
screen of mesh size 40. Maintained the temperature of gelatin mass storage vessel between
2.4 Slurry of Ferric oxide yellow was made in glycerin and purified water. The slurry was passed
through colloid mill for 15-20 minutes. Rinsed the colloid mill with the remaining amount of
purified water for three times. ·
2.5 Added the bulk of step 2.4 to the gelatin mass of step 2.3 in gelatin color mixing assembly for
15-30 minutes at 15-20 RPM at 50-60°C. Apply 600-700 mm Hg and remove entrapped air
bubble while stirring the gelatin mass at low speed. Store the gelatin mass at 55-60°C.
Maintain the temperature of the gelatin storage tank at 55-60°C till encapsulation.
3. Encapsulation, drying and polishing.
3.1 Collected the capsules in drying trays and spread them in a single layer and keep these trays
in a trolley in the encapsulation room until transferring them to drying area. The capsules
. I
3.2
were shuffled at regular interval of2-3 hours.
Transferred the trolley containing drying trays with capsules into the drying area for drying
at 48-72 hours. During drying, the capsules were shuffled at regular intervals till complete
drying.
Dried capsules of step 3.2 were passed through polishing Pan/Tumble dried fitted with lint free
cloth/suitable oil absorbent cloth and rotated for 15-20 minutes.
Example 3
Formulation of Cholecalciferol Syrup (liquid dosage form)
Page 11 of 16
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Each 5 mL of syrup contains: Cholecalciferol (Vitamin 03) ........................... 240000 IU
Table 3: Composition of Cholecalciferol Syrup(liguid dosage form)
Ingredients Qty mg/Sml
Vitamin 03 6.0
Miglyol 812N 21.43
Cremophor RH 40 I 00.525
Propylene glycol 21.43
Tocopherol 0.015
Neosorb P60W 2.5
Sucralose 0.0025
Sodium Methyl Paraben 0.02
Sodium propyl Paraben 0.004
Sodium Benzoate 0.025
Sodium Chloride 0.0025
Water q.s 5 mL
Example 4
In-vitro Drug Release Study
DrugRelease Study_was carried out in Dissolution Apparatus Type II. 500 mL of0.3% SLS in
Purified water was used as a dissolution medium. Dissolution media was stirred at 75 rpm .
. It can be observed from the dissolution profile in Figure 1 that complete drug was released within
an hour. This inferred immediate release ofthe drug.
Example 5
Comparison of in vivo Pharmacokinetic profile of Vitamin DJ PBL formulation (Liquid oral
formulation) and Marketed Formulation (Liquid oral formulation)
Materials
Animal: New Zealand Rabbits
Groups:
Page 12 of 16
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•
1. The I iqu id formulation of the present invention (n=3) (Batch No. 2188/078) (I, 00,000 I U/5 ml)
2. Marketed formulation (n=3) (Depura; Batch No. S2248) (60,000 IU/5 ml)
Dose: I, 00, 000 IU/rabbit
Route of Administration: Per Oral (p.o.)
Total number of animals: 3 rabbits/batch
Duration of Study: I month
Sampling Intervals: 0, 1, 2, 4, 8 hours on day 1 and on day 2, 3, 7, 14, 21 and 28.
Experimental Procedure:
Overnight fasted rabbits were administered with appropriate drug formulation.
The liquid formulation of the present invention (Batch No 2188/078) was administered at a dose of
1, 00,000 IU orally as 5 ml/rabbit. Marketed reference formulation (Batch No S2248) was
administered at a dose of 1 ,00,000 IU orally as 8.33 ml/rabbit. The blood samples were withdrawn
at regular intervals as mentioned above from ear marginal vein. The blood samples collected in
plain tube to collect serum. The concentration of 25(0H) vitamin 03 in the serum was estimated
by ELISA (Kit method).
'
Table 4: Comparison of in vivo Pharmacokinetic profile of Vitamin 03 (PBL formulation) and
Marketed Formulation (Liquid oral formulation)
Group AUC (0-28) (ng*hr/ml) Cmax (ng/ml) Tmax (hr)
Panacea liquid formulation
179382.836 708.27 72
(Batch no. 2188/078)
Marketed Liquid formulation
152273.408 562.1 72
(Depura; S2248)
The liquid formulation of the present invention (Batch no. 2188/078) administered as 5ml
(contains 1,00,000 IU/5ml) has shown AUC(0-28) 179382.84 & Cmax 708.27 ng/ml whereas
marketed liquid formulation (Depura; S2248) administ~red as 8.33 ml (contains 60,000 IU/5ml)
has shown AUC(0-28) 152273.40 & Cmax 562.1 ng/ml.
Page 13 of 16
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The results showed that feed the same active ingredient administered a dose of the nano type
emulsion in comparison with marketed product OEPURA, The liquid formulation of the present
invention showed a more superior performance. The results ofthe PK study are shown in Figure
2. As can be seen in this Figure 2 and it has been found that the compositions of the present
invention permit the preparation of semi-solid and liquid compositions containing an active
vitamin 03 compound in sufficiently high concentration to permit, e.g., convenient oral
administration, while at the same time achieving improved pharmacokinetic parameters for the
active vitamin 03 compound. For example, as compared to OEPURA, The liquid formulation of
the present invention exhibit a Cnax that is at least 1.2 times greater than the Cmax that is observed
with OEPURA, However T max was observed with present invention is similar with observed T max
ofOEPURA.
So we can conclude that liquid formulation of the present· invention has shown better
pharmacokinetic profile as compared to the marketed formulation.
Example 6:
Comparison of in vivo Pharmacokinetic profile of Vitamin 03 PBL formulation (solid Oral) and
Marketed Formulation (Solid oral and Injectable formulation)
Materials
Animal:
Groups:
Dose: 1, 00,000 IU approx
New Zealand Rabbits
1. The capsule formulation of the present invention (n=3) (Batch No.
2150/030A) (1, 00, 000 IU/Cap)
2. Mktd formulation (n=3) (Batch No. UPSG-17055) (60,000 IU/Cap)
3. Mktd formulation (n=3) (Batch No. JACA-7002) (3,00,000 IU/ml)
Route of Administration: Per Oral (p.o.) or Intramuscular (i.m.)
Total number of animals: 3 rabbits/batch
Duration of Study: 1 month
Sam piing Intervals: 0, 1, 2, 8 hours on day 1 and on day 3, 7, 14, 21 and 28.
~ nr~uT 1?-B2-202l 0 T..il:"l-•~ -•• ·lb. v....l n .11- - -
Page 14 of 16
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Experimental Procedure:
Overnight fasted rabbits were administered with appropriate drug. The capsule formulation ofthe
present invention (Batch No. 2150/030A) was administered orally as I, 00,000 IU (I
capsule/rabbit); reference formulation (Batch No. UPSG-17055) was administered orally as
I ,20,000 IU (2 capsule/rabbit) and by intramuscular route (Batch No. JACA-7002) as I ,00,000 IU
(0.33ml/rabbit). The blood samples were withdrawn at regular intervals as mentioned above from
ear marginal vein. The blood samples collected in in plain tube to collect serum. Concentration of
25 (OH) vitamin 03 in serum was estimated by ELISA (Kit method).
The capsule formulation of the present invention (Batch No. 2150/030A-IOO,OOO IU/Cap)
administered as I, 00,000 IU (I Capsule/rabbit) has shown Cmax of 569.35 ng/ml whereas
marketed capsule formulation (Batch No. UPSG-17055-60,000 IU/Cap) administered as I ,20,000
IU (2 Capsules/rabbit) has shown Cmax of 635.0 ng/ml. And marketed injectable formulation
•
(Batch No. JACA-7002-3, 00,000 IU/ml) injected as I ,00,000 IU (0.33 ml/rabbit) has shown
different release profile (i.e. release is less as compared to oral formulation but showing sustained
release up to 28 days). So it can be concluded that both capsule formulation of the present invention·
and reference has shown similar release profile.
Table 5: Comparison of in vivo Pharmacokinetic profile of Vitamin 03 ofPBL formulationf(oral)
vs. Marketed formulations (oral & injectable)
Time point
Group 0 1 8
hour- hour- 2hour- hour- day
(ng/ml) day1 day 1 day 1 day 1 day 3 day 7 day 14 day 21 28
Panacea Average 0.60 3.17 9.23 35.48 569.35 481.87 334.90 120.10 49.83
formulation so 0.10 2.36 10.04 17.51 211.10 187.45 187.67 84.65 31.73
Marketed Average 0.60 5.48 6.14 21.24 635.00 566.33 295.67 146.67 53.17
capsule
formulation so 0.07 0.50 6.94 13.26 171.57 149.75 67.00 51.05 42.35
Average 1.13 4.11 3.83 2.53 11.37 12.63 34.73 57.68 83.39
Page 15 of 16
Marketed
Injectable
formulation so I. 15 1.00 1.12 1.46 1.33 2.23 14.83 28.25 71.27

We claim:

A stable pharmaceutical nano-emulsion composition compnsmg: a) vitamin 03
compound; b) lipophilic phase; c) hydrophilic phase; and/or d) surfactant; and e)
pharmaceutically acceptable excipients.
2. The nano-emulsion composition as claimed in claim I, wherein vitamin 03 is in the form
of nano-oil globules dispersed in aqueous phase and characterized in that the 50 percent
{ d50) of globule (s) of said nano-emulsion have a size less than I OOnm throughout the
shelf-life.
3. The nano-emulsion composition as claimed in claim I, wherein the amount of the vitamin
03 ranges between 0.0 I to 20% w/v, preferably between 0.1 to I 0% w/v.
4. The nano-emulsion composition as claimed in claim I, wherein the lipophilic phase
component is selected from the group consisting of mono, di or triglycerides, preferably
triglycerides and combinations thereof.
5. The nano-emulsion composition as claimed m claim 4, wherein the amount of the
triglyceride ranges between 5 to 30 % w/v, preferably between I 0 to 20 % w/v.
6. The nano-emulsion composition as claimed in claim I, wherein the hydrophilic phase
component is selected from the group consisting ofCt-s alkyl or tetrahydrofurfuryl di or
partial-ether of a low molecular weight mono or poly-oxy-alkane diol, preferably polyoxy-
alkane diol.
7. The nano-emulsion composition as claimed in claim 6, wherein the amount of the polyoxy-
alkane diol ranges between 5 to 30% w/v, preferably between 10 to 20% w/v.
8. The nano-emulsion composition as claimed in claim 1, wherein the surfactant is selected
from the group consisting of polyoxyalkylenes dextrans, fatty acid esters of saccharose,
fatty alcohol ethers of oligoglucosides, fatty acid esters of glycerol, polyoxyethylene
compounds, polyethoxylated fatty acid esters of sorbitan, fatty acid esters of polyethylene
-Q)
C)
Ill
D..
Q) -1-- N
E....
0
LL.
(';
"-:":1'"'
(0
0
0 ..........
0
N
0
~ en
N
-.~.... .....
N
0
N
I .c
Q)
LL.
I
.N... .
oxide, fatty alcohol ethers of polyethylene oxide, alkylphenol ethers ofpotyethylene oxide;
polyoxyethylene-polyoxypropylene block copolymers, ethoxylated fatty oils and
combinations thereof.
9. The nano-emulsion composition as claimed m claim 8, wherein the amount of the
-
surfactant,ranges between 30 to 80% w/v, preferably between 40 to 70% w/v.
10. A stable pharmaceutical nano-emulsion composition as claimed in claim I comprising: a)
vitamin 03 compound in an amount ofO.OI to 20% w/v, preferably, 0.1 to 10% w/v; b)
at least one· lipophilic phase component in an amount of 5 to 30% w/v, preferably I 0 to 20
% w/v; c) at least one hydrophilic phase component in an amount of 5 to 30 % w/v,
preferably I 0 to 20% w/v; d) at least one non-ionic surfactant in an amount of 30 to 80%
w/v, preferably, 40 to 70% w/v; and e) at least one pharmaceutically acceptable excipient.