The present disclosure relates generally to pharmaceutical formulations. More specifically, the disclosure is directed to an intranasal nanoemulgel formulation for brain delivery of active component berberine hydrochloride. The present disclosure also provides a process for manufacturing the formulation.
BACKGROUND OF THE INVENTION
[0002] Background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art. [0003] The practical limitations associated with most of the drug delivery systems associated with delivering the drug to the brain, involves poor delivery because of low permeability across the blood brain barrier. Blood brain barrier is a network of blood cells and vessels that forms a permeable layer that selectively permits solutes from the blood to enter the fluid of the central nervous system. This prevents harmful pathogens and toxins from entering the brain but also ends up blocking some of the essential drugs. Apart from this the known systems associated with brain delivery, lack in mode of application, biodegradability, or irregularity in drug release behaviour. Some formulations involve nasal delivery of liquid dosage forms, which are often limited due to its short residence time as they are quickly cleared off from the nasal cavity. Possible options for drug delivery systems include creams, solutions, and ointments prepared by conventional mixing and chemical cross-linking methods. However, they have their limitation of applicability, flow stability and uncontrolled drug release which further causes irregularity in drug release and low permeability across the blood brain barrier.
[0004] Berberine hydrochloride is an anti-tumor, anti-microbial, anti-diabetic, and anti-inflammatory compound. It also has multiple neuro-protective and pharmacological effects in the brain but it is riddled by poor solubility and low permeability. Therefore there is a need in the art to develop formulations of

berberine hydrochloride that improve drug delivery in the brain by effectively crossing the blood brain barrier.
OBJECTS OF THE INVENTION
[0005] An object of the present disclosure is to provide an intranasal nanoemulgel
formulation for brain delivery of berberine hydrochloride.
[0006] An object of the present disclosure is to provide a formulation for
berberine hydrochloride that enhances its permeability across the blood-brain
barrier.
[0007] Another object of the present disclosure is to provide a formulation that is
biodegradable, has a regular release of drug and has long residence time in the
nasal cavity.
[0008] Yet another object of the present disclosure is to provide a process of
manufacturing an intranasal nanoemulgel formulation for brain delivery of
berberine hydrochloride.
SUMMARY OF THE INVENTION
[0009] This summary is provided to introduce a selection of concepts in a simplified form that are further described below in Detailed Description section. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter.
[0010] Aspects of the present disclosure provide an intranasal nanoemulgel formulation comprising berberine hydrochloride that is topically applied for brain delivery of the drug.
[0011] In an aspect, the present disclosure provides an intranasal nanoemulgel formulation for brain delivery of berberine hydrochloride, wherein the formulation comprises a carbomer and a nanoemulsion composition comprising berberine hydrochloride, calendula oil, a polysorbate and a polyoxyethylene-polyoxypropylene block copolymer.

[0012] In an embodiment, the formulation comprises berberine hydrochloride in a range of about 0.1% w/v to about 0.5%w/v of the formulation. [0013] In an embodiment, the formulation comprises calendula oil in a range of about 0.1% v/v to about l%v/v of the formulation.
[0014] In an embodiment, the polysorbate may be selected from polyoxyethylene (20) sorbitan monolaurate (polysorbate 20), polyoxyethylene (20) sorbitan monopalmitate (polysorbate 40), polyoxyethylene (20) sorbitan monostearate (polysorbate 60), polyoxyethylene (20) sorbitan monooleate (polysorbate 80), or combinations thereof.
[0015] In an embodiment, the formulation comprises polysorbate in a range of about P/o v/v to about 5%v/v of the formulation.
[0016] In an embodiment, the formulation comprises polyoxyethylene-polyoxypropylene block copolymer in a range of about 0.1% w/v to about l%w/v of the formul ati on.
[0017] In an embodiment, the polysorbate and polyoxyethylene-polyoxypropylene block copolymer may be present in a ratio range of about 1:1 to about 1:5.
[0018] In an embodiment, the carbomer may be a carbopol present in the formulation in a range of about 0.5% w/v to about 3%w/v of the formulation. [0019] In an embodiment, the formulation further comprises at least one co-surfactant. In an embodiment, the co-surfactant may be ethanol, methanol, or their combination. In a preferred embodiment, the co-surfactant may be ethanol. [0020] In an embodiment, the formulation comprises co-surfactant in a range of about 0.1%> v/v to about l%v/v of the formulation.
[0021] In an embodiment, the formulation further comprises a solvent selected from distilled water, dimeralized water or water. In an embodiment, the solvent may be distilled water present in the formulation in a range of about 90% v/v to about 95%v/v of the formulation.
[0022] In an aspect, the present disclosure provides a process of manufacturing an intranasal nanoemulgel formulation for brain delivery of berberine hydrochloride, wherein the process comprises the steps of: (a) dissolving berberine hydrochloride

in calendula oil to give an organic phase; (b) dissolving a polyoxyethylene-polyoxypropylene block copolymer in ethanol and mixing the organic phase into it; (c) preparing an aqueous phase by dissolving a polysorbate in distilled water followed by adding the solution of (b) into the aqueous phase to give an emulsion; (d) ultrasonicating the emulsion to give an oil in water nanoemulsion; and (e) blending a carbomer with the nanoemulsion to give the nanoemulgel formulation. [0023] Other aspects of the invention will be set forth in the description which follows, and in part will be apparent from the description, or may be learnt by the practice of the invention.
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] The following drawings form part of the present specification and are included to further illustrate aspects of the present disclosure. The disclosure may be better understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein.
[0025] Figure 1(A) provides a photographic image of a nanoemulsion (white) without berberine hydrochloride (blank) and a nanoemulgel formulation (orange) comprising berberine hydrochloride as per an embodiment of the present disclosure. Figures 1(B), 1(C) and 1(D) provide Transmission Electron Microscopy (TEM) images of the nanoemulgel formulation, as per an embodiment of the present disclosure, showing the morphology and shape of the prepared nanoemulgel formulation.
[0026] Figure 2 graphically represents the percentage drug release (%) with time (h) or drug release profile of different formulations prepared as per an embodiment of the present disclosure.
[0027] Figure 3 graphically represents the ex vivo drug permeation study giving the percentage drug permeation (%) with time (h) of different formulations prepared as per an embodiment of the present disclosure.

DETAILED DESCRIPTION OF THE INVENTION
[0028] The following is a detailed description of embodiments of the disclosure. The embodiments are in such detail as to clearly communicate the disclosure. However, the amount of detail offered is not intended to limit the anticipated variations of embodiments; on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present disclosure as defined by the appended claims.
[0029] All publications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Where a definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply. [0030] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.
[0031] In some embodiments, numbers have been used for quantifying weights, percentages, ratios, and so forth, to describe and claim certain embodiments of the invention and are to be understood as being modified in some instances by the term "about." Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the invention

are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable. The numerical values presented in some embodiments of the invention may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements. [0032] Various terms as used herein are shown below. To the extent a term used in a claim is not defined below, it should be given the broadest definition persons in the pertinent art have given that term as reflected in printed publications and issued patents at the time of filing.
[0033] As used in the description herein and throughout the claims that follow, the meaning of "a," "an," and "the" includes plural reference unless the context clearly dictates otherwise. Also, as used in the description herein, the meaning of "in" includes "in" and "on" unless the context clearly dictates otherwise. [0034] Unless the context requires otherwise, throughout the specification which follow, the word "comprise" and variations thereof, such as, "comprises" and "comprising" are to be construed in an open, inclusive sense that is as "including, but not limited to."
[0035] The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. [0036] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. "such as") provided with respect to certain embodiments herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention. [0037] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member can be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group can be

included in, or deleted from, a group for reasons of convenience and/or
patentability. When any such inclusion or deletion occurs, the specification is
herein deemed to contain the group as modified.
[0038] The description that follows, and the embodiments described therein, is
provided by way of illustration of an example, or examples, of particular
embodiments of the principles and aspects of the present disclosure. These
examples are provided for the purposes of explanation, and not of limitation, of
those principles and of the disclosure.
[0039] It should also be appreciated that the present disclosure can be
implemented in numerous ways, including as a system, a method or a device. In
this specification, these implementations, or any other form that the invention may
take, may be referred to as processes. In general, the order of the steps of the
disclosed processes may be altered within the scope of the invention.
[0040] The headings and abstract of the invention provided herein are for
convenience only and do not interpret the scope or meaning of the embodiments.
[0041] The following discussion provides many example embodiments of the
inventive subject matter. Although each embodiment represents a single
combination of inventive elements, the inventive subject matter is considered to
include all possible combinations of the disclosed elements. Thus if one
embodiment comprises elements A, B, and C, and a second embodiment
comprises elements B and D, then the inventive subject matter is also considered
to include other remaining combinations of A, B, C, or D, even if not explicitly
disclosed.
[0042] As described herein, the term 'therapeutically effective amount' refers to
the amount of the formulation required to bring about a change or improvement in
a subject without side effects or overdosing.
[0043] The term, "subject" as used herein refers to an animal, preferably a
mammal, and most preferably a human. The term "mammal" used herein refers to
warm-blooded vertebrate animals of the class 'mammalia' , including humans,
characterized by a covering of hair on the skin and, in the female, milk-producing

mammary glands for nourishing the young, the term mammal includes animals
such as cat, dog, rabbit, bear, fox, wolf, monkey, deer, mouse, pig and human.
[0044] As used herein, the term 'nanoemulgel' refers to a gel formulation of a
nanoemulsion composition comprising berberine hydrochloride, wherein the gel
formulation comprises nanoparticles.
[0045] Aspects of the present disclosure provide an intranasal formulation that
improves the brain delivery of the drug berberine hydrochloride via olfactory
region.
[0046] In an embodiment, the present disclosure provides an intranasal
nanoemulgel formulation for brain delivery of berberine hydrochloride, wherein
the formulation comprises a gelling agent and a nanoemulsion composition
comprising berberine hydrochloride.
[0047] In an embodiment, the present disclosure provides an intranasal
nanoemulgel formulation for brain delivery of berberine hydrochloride, wherein
the formulation comprises a carbomer and a nanoemulsion composition
comprising berberine hydrochloride, calendula oil, a polysorbate and a
polyoxyethylene-polyoxypropylene block copolymer.
[0048] In an embodiment, the formulation comprises berberine hydrochloride in a
range of about 0.1% w/v to about 0.5%w/v of the formulation. In a preferred
embodiment, the formulation comprises berberine hydrochloride in a range of
about 0.1%) w/v to about 0.3%w/v of the formulation.
[0049] In an embodiment, the formulation comprises calendula oil in a range of
about 0.1%) v/v to about l%v/v of the formulation. In a preferred embodiment, the
formulation comprises calendula oil in a range of about 0.1%> v/v to about
0.5%v/v of the formulation.
[0050] In an embodiment, the polysorbate may be selected from polyoxyethylene
(20) sorbitan monolaurate (polysorbate 20), polyoxyethylene (20) sorbitan
monopalmitate (polysorbate 40), polyoxyethylene (20) sorbitan monostearate
(polysorbate 60), polyoxyethylene (20) sorbitan monooleate (polysorbate 80), or
combinations thereof. In a preferred embodiment, the polysorbate is
polyoxyethylene (20) sorbitan monooleate (polysorbate 80).

[0051] In an embodiment, the formulation comprises polysorbate in a range of about l%v/v to about 5%v/v of the formulation. In a preferred embodiment, the formulation comprises polysorbate in a range of about 1% v/v to about 2%v/v of the formulation.
[0052] In an embodiment, the formulation comprises polyoxyethylene-polyoxypropylene block copolymer in a range of about 0.1% w/v to about l%w/v of the formulation. In a preferred embodiment, the formulation comprises the copolymer in a range of about 0.1% w/v to about 0.5%w/v of the formulation. [0053] The polysorbate and polyoxyethylene-polyoxypropylene block copolymer act as surfactant to stabilize the nanoemulsion. Further, the surfactants are biocompatible and being P-gp substrates increases the transfer of berberine hydrochloride across the blood-brain barrier. Both the surfactants not only possess excellent biocompatibility and water solubility but also offer tunable drug release as well as high permeation of berberine hydrochloride across the nasal mucosa. [0054] In an embodiment, the polysorbate and polyoxyethylene-polyoxypropylene block copolymer may be present in a ratio range of about 1:1 to about 1:5. In a preferred embodiment, the polysorbate and polyoxyethylene-polyoxypropylene block copolymer may be present in a ratio of about 1:2. [0055] In this disclosure, nano-gelation has been achieved by mixing nanoemulsion (NE) in a gelling agent - carbomer which in-turn forms into a nanoemulgel. The nanoemulsion is an oil in water emulsion. [0056] In an embodiment, the carbomer may be a carbopol.
[0057] In an embodiment, the formulation comprises carbomer in a range of about 0.5% w/v to about 3%w/v of the formulation. In a preferred embodiment, the formulation comprises carbomer in a range of about 0.5% w/v to about 1.5%w/v of the formul ati on.
[0058] In an embodiment, the formulation further comprises at least one co-surfactant. In an embodiment, the co-surfactant may be ethanol, methanol, or their combination. In a preferred embodiment, the co-surfactant may be ethanol. [0059] In an embodiment, the formulation comprises co-surfactant in a range of about 0.1%) v/v to about 1%>V/V of the formulation.

[0060] In an embodiment, the formulation further comprises a solvent selected from distilled water, demineralized water or water. In an embodiment, the solvent may be distilled water present in the formulation in a range of about 90% v/v to about 95%v/v of the formulation.
[0061] In an embodiment, the formulation may also comprise one or more additive(s). The additive(s) may be selected from preservatives, pH adjusting agent, coloring agent or dyes, fragrance, or combinations thereof. However, a person skilled in the art would understand that any other 'additive' may be added to the formulation without going beyond the spirit and scope of the present disclosure.
[0062] In an embodiment, the preservative may be selected from sorbic acid, parabens such as methyl or propyl parabens, or combinations thereof. In an embodiment, the pH adjusting agent may be triethanolamine, acetic acid or combinations thereof. In an embodiment, the coloring agent may be any well-known organic or inorganic dyes, natural or synthetic dyes or combinations thereof. In an embodiment, the fragrance may be any fragrant compound or natural extract that is suitable for topical application including menthol, rose or neem extract.
[0063] The formulation synergistcally provides increased permeation across the blood brain barrier and has longer residence time in the nasal mucosa for the drug. This avoids the hepatic metabolism of the drug and consequent low bioavailability. The formulation comes in contact with the nasal mucosal membrane and dissolves to release the drug.
[0064] In an embodiment, the formulation provides a regular drug release ensuring uniformity of dose. In some embodiments, the formulation may have uniformity of dosage for a period of up to 16 h.
[0065] The formulation is a gel based formulation making it easy to apply and need not be removed after application as it is completely biodegradable. [0066] The formulation is cost-effective and has higher patient compliance compared to orally administered drug delivery systems and nasal liquid dosage forms, as it can be applied by the patient himself/herself

[0067] The formulation is a synergistic formulation that is stable, has high nasal
retention, and high permeation across the blood brain barrier. The formulation is
more effective than marketed solutions, ointments and creams for nasal delivery.
[0068] The formulation is stable with zeta potentials generally in the range of ±30
mV. The formulation also does not lead to separation of the aqueous and organic
phase, creaming or cracking.
[0069] In an embodiment, the present disclosure provides a medicament
comprising the intranasal nanoemulgel formulation for topical application.
[0070] In some embodiments, the medicament may comprise one or more other
active agent(s) for brain delivery.
[0071] In another embodiment, the present disclosure provides use of an
intranasal nanoemulgel formulation of berberine hydrochloride as recited above,
for effective neuroprotective and pharmacological effect in the brain.
[0072] In an embodiment, the formulation may be used for treating, ameliorating,
or preventing Alzheimer's disease, Parkinson's diseases and other related
disorders of the brain. The formulation effectively delivers berberine
hydrochloride to the brain through the nasal route.
[0073] In an embodiment, the formulation may be used for treating, ameliorating,
or preventing symptoms associated with Alzheimer's disease, Parkinson's
diseases and other related disorders of the brain.
[0074] In an embodiment, the formulation is placed in nasal cavity or delivered
through nasal route. The formulation increases nasal retention time and
subsequently releases the drug leading to its rapid absorption across the blood
brain barrier.
[0075] In an embodiment, the present disclosure provides a process of
manufacturing an intranasal nanoemulgel formulation using: (a) ultrasonication
for preparation of a nanoemulsion and (b) homogenization for converting the
nanoemulsion into nanoemulgel.
[0076] In yet another embodiment, the present disclosure provides a process of
manufacturing an intranasal nanoemulgel formulation for brain delivery of
berberine hydrochloride, wherein the process comprises the steps of: (a)

dissolving berberine hydrochloride in calendula oil to give an organic phase; (b)
dissolving a polyoxyethylene-polyoxypropylene block copolymer in ethanol and
mixing the organic phase into it; (c) preparing an aqueous phase by dissolving a
polysorbate in water followed by adding the solution of (b) into the aqueous phase
to give an emulsion; (d) ultrasonicating the emulsion to give an oil in water
nanoemulsion; and (e) blending a carbomer with the nanoemulsion to give the
nanoemulgel formulation.
[0077] The process is cost-effective and industrially scalable.
[0078] In another embodiment, the present disclosure provides a method of
enhancing neuroprotective effect of berberine hydrochloride in a subject, wherein
the method comprises the steps of administering a therapeutically effective
amount of the formulation as recited above.
[0079] The therapeutically effective amount may be decided by a trained
physician or doctor based on the medical history, weight, age, gender, and other
characteristics of the subject.
[0080] While the foregoing describes various embodiments of the disclosure,
other and further embodiments of the disclosure may be devised without departing
from the basic scope thereof. The scope of the invention is determined by the
claims that follow. The invention is not limited to the described embodiments,
versions or examples, which are included to enable a person having ordinary skill
in the art to make and use the invention when combined with information and
knowledge available to the person having ordinary skill in the art.
EXAMPLES
[0081] The present invention is further explained in the form of following
examples. However, it is to be understood that the following examples are merely
illustrative and are not to be taken as limitations upon the scope of the invention.
[0082] EXAMPLE 1: Preparation of nanoemulgel formulation
[0083] Berberine hydrochloride Nanoemulgel formulations were prepared as per
the formulation provided in Table No. 1.10 mg of berberine hydrochloride was
dissolved in 0.5 mL of calendula oil to give an oil phase. This was followed by
mixing of the oil phase in a mixture of 0.5 mL pluronic® F68 and 1 mL ethanol.

The above solution was added drop-wise into water phase containing 1 mL of Tween® 80 as hydrophilic surfactant. Oil in water nanoemulsion was formed using ultrasonication process for a time period of 10 minutes (30 seconds sonication was on and for 30 seconds sonication was off). lOmL of the prepared nanoemulsion was taken for the preparation of the Nanoemulgel formulation. Nanoemulgel was prepared by mixing varying concentrations of the gelling agent i.e. Carbopol® 934 (0.5%w/v to 3%w/v) with the nanoemulsion and the mixture was stirred for 24 hrs for complete mixing of nanoemulsion and gelling agent to give the nanoemulgel formulation.
[0084] One comparative formulation was also prepared - Blank nanoemulsion in which the drug was dissolved in distilled water. The permeation of this was compared with that of drug encapsulated in nanoemulgels.
[0085] The physico-chemical parameters of the formulation were determined by the procedure provided below and the results are presented in Table 1. [0086] Particle size and zeta potential studies: Droplet size and zeta potential of prepared nanoemulsions were obtained using Nanotrac Wave Zetasizer (Microtrac, USA). Samples were diluted with de-ionized water before measurement. Zeta potential of the samples was also measured as an indicator of physical stability within the media.
[0087] Drug encapsulation efficiency of nanoemulsions: The encapsulation efficiency (EE) values of the nanoemulgel with berberine hydrochloride were determined using UV spectrophotometer at Xmax (348 nm). A sample of each formulation (nanoemulgel) was diluted in methanol and amount of berberine encapsulated was determined by measuring the non-encapsulated berberine in the aqueous phase after ultrafiltration/centrifugation. The berberine encapsulation efficiency was calculated from the difference between the concentrations of the total from that of the free berberine hydrochloride in distilled water.

Table 1: Composition of nanoemulgel loaded with berberine
hydrochloride ~S! Cod Oil Drug Pluron Twee Carbop Distille Particl Zeta (%"
No e (%v/ (%w/ icF68 n80 ol (% d e size Potenti EE
v) v) (%w/v) (%v/ w/v) water (nm) al (mV) )
v) (ml)

1 Fl - 0.1 - 10 - -
2 F2 0.2 0.1 0.2 0.4 0.5 10 112 +4.3 78
3 F3 0.3 0.1 0.3 0.6 1 10 145 +5.8 82
4 F4 0.4 0.1 0.4 0.8 1.5 10 198 +6.7 89
10 236 +8.2 90
6 F6 0.4 0.1 0.6 0.8 2.5 10 245 +9.3 86
7 F7 0.3 0.1 1.0 0.6 3 10 202 +11.2
[0088] Figure 1(A) provides an image of a blank nanoemulsion (white) (no gelling agent and drug) and the nanoemulgel formulation comprising berberine hydrochloride (orange) of the present disclosure. Figures 1(B), 1(C) and 1(D) provide the TEM images of the nanoemulgel formulation showing the morphology and shape of the prepared nanoemulgel. [0089] EXAMPLE 2: Drug release profile
[0090] In-vitro drug release study from formulations of Table 1, was performed using dialysis bag method. An appropriate amount of formulation was suspended in the dialysis bag (Mol wt. 12000-14000) and immersed into the release medium at 37