METHOD FOR TREATMENT OF OSTEOARTHRITIS USING COMBINATION OF IBUPROFEN AND GLUCOSAMINE SULPHATE AND ITS COMPOSITION

Abstract

ABSTRACT METHOD FOR TREATMENT OF OSTEOARTHRITIS USING COMBINATION OF IBUPROFEN AND GLUCOSAMINE SULPHATE AND ITS COMPOSITION The present invention provides a pharmaceutically acceptable dosage form of ibuprofen-glucosamine sulphate nanocrystals for the treatment of joint pain and inflammation in osteoarthritis with negligible side effects in addition to improved antinociceptive and disease modifying potential.

Specification

Description:FORM 2
THE PATENTS ACT, 1970
(39 OF 1970)
&
THE PATENTS RULES, 2003
COMPLETE SPECIFICATION
(See section 10; rule 13)



Title: METHOD FOR TREATMENT OF OSTEOARTHRITIS USING COMBINATION OF IBUPROFEN AND GLUCOSAMINE SULPHATE AND ITS COMPOSITION


APPLICANT DETAILS:
(a) NAME: NATIONAL INSTITUTE OF PHARMACEUTICAL EDUCATION
AND RESEARCH (NIPER), HYDERABAD
(b) NATIONALITY: Indian
(c) ADDRESS: Balanagar, Hyderabad, Telangana 500037, India







PREAMBLE TO THE DESCRIPTION:
The following specification particularly describes the nature of this invention and the manner in which it is to be performed.
METHOD FOR TREATMENT OF OSTEOARTHRITIS USING COMBINATION OF IBUPROFEN AND GLUCOSAMINE SULPHATE AND ITS COMPOSITION
Field of Invention:
The present invention relates to a composition and method for the management of joint pain and inflammation in osteoarthritis.

Background of the Invention:
The following background discussion 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 expressly or implicitly referenced is prior art.
Osteoarthritis is a chronic degenerative musculoskeletal disorder characterized by progressive loss of hyaline cartilage followed by subchondral bone remodeling and inflammation. Despite ongoing research in drug discovery, no FDA-approved drugs for osteoarthritis are available. Currently, non-steroidal anti-inflammatory drugs are employed to mitigate the joint pain and inflammation. For example, US7431948B2 discloses a natural formulation of compounds that would modulate inflammation. The formulation would also inhibit expression of COX-2, inhibit synthesis of prostaglandins selectively in target cells, and inhibit inflammatory response selectively in target cells. The compositions containing at least one fraction isolated or derived from hops. Other embodiments relate to combinations of components, including at least one fraction isolated or derived from hops, tryptanthrin and conjugates thereof, rosemary, an extract or compound derived from rosemary, a triterpene species, or a diterpene lactone or derivatives or conjugates thereof.
Further, US9561174B2 discloses a preparation of ibuprofen (2-(4-isobutylphenyl) propionic acid) in the free acid form that is suitable for topical administration. The topical ibuprofen formulation is prepared by dissolving the free acid form of ibuprofen or preparing a homogeneous suspension of the free acid form of ibuprofen, in the presence of a pharmaceutically acceptable solvent so as to produce a topical drug formulation compatible with the penetration of 2-(4-isobutylphenyl) propionic acid through the skin tissue. Topical formulations of ibuprofen can be based on a pharmaceutically acceptable solvent such as, e.g., a pyrrolidone solvent or dimethylacetamide.
In another documents, US7759317B2 discloses pharmaceutical compositions containing certain flavonoid glycoside-type compounds that have been shown in the present invention to be therapeutically useful, including analgesics and anti-inflammatories for the treatment and management of pain and inflammatory conditions. Methods for the therapeutic uses of such compounds and pharmaceutical compositions is also provided.
The above-mentioned documents disclose ibuprofen as compound which only provides symptomatic relief to the joint pain. The long-term use of the ibuprofen has certain side effects such as swelling of face and lower extremities, severe stomach pain and gastrointestinal bleeding. There is need of the improved compound and method to minimize the side effects while improving its antinociceptive and disease modifying potential.
The present invention provides a pharmaceutically acceptable dosage form of ibuprofen-glucosamine sulphate nanocrystals for the treatment of joint pain and inflammation in osteoarthritis with negligible side effects in addition to improved antinociceptive and disease modifying potential.

Object(s) of the present invention:
The primary objective of the present invention is to overcome the drawback associated with prior art.
An object of the present invention is to provide a pharmaceutically acceptable dosage form of ibuprofen-glucosamine sulphate nanocrystals for the treatment of joint pain and inflammation in osteoarthritis with negligible side effects in addition to improved antinociceptive and disease modifying potential.

Summary of the Invention:
In an aspect the present invention provides a composition for management of joint pain and inflammation in osteoarthritis comprising:
a) ibuprofen present in amount of 50-600 mg;
b) methanol present in amount of 5 mL, wherein the ibuprofen in amount of 50-600 mg is mixed with the methanol in amount of 5 mL to form an organic phase;
c) glucosamine sulphate present in amount of 5-10%; and
d) a hydrophilic non-ionic surfactant, wherein an aqueous phase is prepared by dissolving the glucosamine sulphate present in amount of 5-10% in 20 mL of distilled water;
wherein the organic phase is dropwise injected into the aqueous phase under continuous stirring followed by probe sonication in addition to evaporation of methanol to form nanocrystals of the ibuprofen and the glucosamine sulphate.
In an embodiment, nanocrystals of the ibuprofen and the glucosamine sulphate is supplemented with a 1% mannitol and a stored at -80°C and followed by a lyophilization to obtain powdered form of the ibuprofen and glucosamine sulphate nanocrystals.
In an aspect the present invention provides a method for synthesis of a composition for management of joint pain and inflammation in osteoarthritis comprising steps of:
a) dispersing ibuprofen in amount of 50-600 mg in methanol to form an organic phase;
b) dissolving glucosamine sulphate in amount of 5-10% with a hydrophilic non-ionic surfactant in 20 mL distilled water to form an aqueous phase;
c) injecting the organic phase obtained at step (a) dropwise injected into the aqueous phase obtained at step (b) under continuous stirring condition with evaporation of methanol;
d) agitating particles solution obtained at step (c) by a probe sonication for duration of a 5 min with a 10 sec on and a 5 sec off cycle to obtain nanocrystals; and
e) supplementing nanocrystals obtained at step (d) with 1% mannitol and storing at temperature of -80°C followed by lyophilization to obtain powdered form of nanocrystals.
In an embodiment, the powdered form of nanocrystals of the ibuprofen and the glucosamine sulphate is synthesized into a gel form by steps of:
a) dispersing Ibuprofen-glucosamine sulphate nanocrystals in 2% w/v Carbopol Ultrez 10-NF along with 0.01% methyl paraben and stirring solution overnight followed by pH adjustment from 5 to 6 using triethanolamine.

Brief description of Drawings:
The accompanying drawings, which are incorporated in and constitute a part of this disclosure, illustrate exemplary embodiments and, together with the description, explain the disclosed principles. The reference numbers are used throughout the figures to describe the features and components. Some embodiments of system and/or methods in accordance with embodiments of the present subject matter are now described, by way of example only, and regarding the accompanying figures, in which
Figure 1: illustrates chromatogram for ibuprofen eluted by acetonitrile: orthophosphoric acid (pH~3.5) (75:25 ratio) in C18 column at 222 nm.
Figure 2: illustrates particle size distribution of optimized ibuprofen-glucosamine sulphate nanocrystals (B) Scanning electron microscopy of ibuprofen (C) Scanning electron microscopy of optimized ibuprofen-glucosamine sulphate nanocrystals.
Figure 3: illustrates solubility of ibuprofen and ibuprofen-glucosamine sulphate nanocrystals (IBU-GS-NC) in distilled water.
Figure 4: illustrates viscosity of optimized ibuprofen-glucosamine sulphate nanocrystals loaded gel compared to marketed diclofenac emulgel
Figure 5: illustrates spreadibility of optimized ibuprofen-glucosamine sulphate nanocrystals loaded gel compared to marketed diclofenac emulgel.
Figure 6: illustrates permeation of ibuprofen from optimized ibuprofen-glucosamine sulphate nanocrystals loaded gel and ibuprofen gel across rat skin mounted in Franz-diffusion cell apparatus maintained at 32±2°C.
Figure 7: illustrates thickness (mm) of right knee joint before and post monosodium iodoacetate (3 mg/ 50 µL) induced osteoarthritis followed by various treatments. MIA: Monosodium iodoacetate, IBU gel: Conventional ibuprofen gel, GS gel: Glucosamine sulphate gel; IBU-NC gel: Ibuprofen nanocrystal loaded gel; IBU-GS-NC gel: Ibuprofen-glucosamine sulphate nanocrystals loaded gel; Blank gel: Carbopol Ultrez gel.
Figure 8: illustrates radiographs for (A) normal control, (B) Monosodium iodoacetate-induced osteoarthritis, (C) Monosodium iodoacetate induced osteoarthritis+ Ibuprofen topical gel (IBU Gel), (D) Monosodium iodoacetate induced osteoarthritis + Glucosamine sulphate gel (GS-Gel), (E) Monosodium iodoacetate induced osteoarthritis + Ibuprofen nanocrystals gel (IBU-NC-Gel), (F) Monosodium iodoacetate induced osteoarthritis + Optimized ibuprofen-glucosamine sulphate nanocrystals loaded gel (IBU-GS-NCs-Gel) and (G) Monosodium iodoacetate induced osteoarthritis + Blank carbopol gel treated right knee. (H) Kellgren-Lawrence based scoring system to evaluate the presence and severity of joint space narrowing and osteophytes.
Figure 9: illustrates representative images for (I) safranin O-fast green and (II) toluidine blue stained histological sections. (A) normal control, (B) Monosodium iodoacetate-induced osteoarthritis, (C) Monosodium iodoacetate + ibuprofen gel (IBU-Gel), (D) Monosodium iodoacetate+ glucosamine sulphate gel (GS-Gel), (E) Monosodium iodoacetate+ ibuprofen nanocrystals gel (IBU-NCs), (F) Monosodium iodoacetate + optimized ibuprofen-glucosamine sulphate nanocrystals loaded gel (IBU-GS-NCs and (G) Monosodium iodoacetate + blank carbopol gel treated right knee. (III) The Osteoarthritis Research Society International (OARSI) score based histological evaluation for cartilage degradation assessment
Figure 10: illustrates fold change in expression of (A) COX-2, (B) TNF-α and (C) IL-1β in synovial tissue of normal control, positive control, conventional ibuprofen gel, glucosamine sulphate gel, ibuprofen nanocrystals gel, blank gel and ibuprofen-glucosamine sulphate nanocrystals loaded gel. (D) Representative blots for COX-2, TNF-α, IL-1ß and ß-actin in synovial tissue of rat knee treated with various interventions. MIA: Monosodium iodoacetate, IBU gel: Conventional ibuprofen gel, GS gel: Glucosamine sulphate gel; IBU-NC gel: Ibuprofen nanocrystals loaded gel; IBU-GS-NC gel: Ibuprofen-glucosamine nanocrystals loaded gel; Blank gel: Carbopol Ultrez gel.

Detailed description of the invention:
In the present document, the word "exemplary" is used herein to mean "serving as an example, instance, or illustration." Any embodiment or implementation of the present subject matter described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments.
While the disclosure is susceptible to various modifications and alternative forms, specific embodiment thereof has been shown by way of example, in the drawings and will be described in detail below. It should be understood, however, that it is not intended to limit the disclosure to the specific forms disclosed, but on the contrary, the disclosure is to cover all modifications, equivalents, and alternative falling within the spirit and the scope of the disclosure.
The terms "comprises", "comprising", "includes", or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a setup, device or method that comprises a list of components or steps does not include only those components or steps but may include other components or steps not expressly listed or inherent to such setup or device or method. In other words, one or more elements in a system or apparatus proceeded by "comprises... a" does not, without more constraints, preclude the existence of other elements or additional elements in the system or method.
In an embodiment, the present invention provides a pharmaceutically acceptable dosage form of ibuprofen-glucosamine sulphate nanocrystals for the treatment of joint pain and inflammation in osteoarthritis with negligible side effects in addition to improved antinociceptive and disease modifying potential.
In an aspect the present invention provides a composition for management of joint pain and inflammation in osteoarthritis comprising:
a) ibuprofen present in amount of 50-600 mg;
b) methanol present in amount of 5 mL, wherein the ibuprofen in amount of 50-600 mg is mixed with the methanol in amount of 5 mL to form an organic phase;
c) glucosamine sulphate present in amount of 5-10%; and
d) a hydrophilic non-ionic surfactant, wherein an aqueous phase is prepared by dissolving the glucosamine sulphate present in amount of 5-10% in 20 mL of distilled water;
wherein the organic phase is dropwise injected into the aqueous phase under continuous stirring with evaporation of methanol followed by probe sonication to form nanocrystals of the ibuprofen-glucosamine sulphate.
In an embodiment, the nanocrystals of the ibuprofen and the glucosamine sulphate is supplemented with a 1% mannitol and a stored at -80°C and followed by a lyophilization to obtain powdered form of the ibuprofen and glucosamine sulphate nanocrystals.
In an aspect the present invention provides a method for synthesis of a composition for management of joint pain and inflammation in osteoarthritis comprising steps of:
a) dispersing ibuprofen in amount of 50-600 mg in methanol to form an organic phase;
b) dissolving glucosamine sulphate in amount of 5-10% with a hydrophilic non-ionic surfactant in 20 mL distilled water to form an aqueous phase;
c) injecting the organic phase obtained at step (a) dropwise injected into the aqueous phase obtained at step (b) under continuous stirring condition with evaporation of methanol;
d) agitating particles solution obtained at step (c) by a probe sonication for duration of a 5 min with a 10 sec on and a 5 sec off cycle to obtain nanocrystals; and
e) supplementing nanocrystals obtained at step (d) with 1% mannitol and storing at temperature of -80°C followed by lyophilization to obtain powdered form of nanocrystals.
In an embodiment, the powdered form of nanocrystals of the ibuprofen and the glucosamine sulphate is synthesized into a gel form by steps of:
b) dispersing 2% w/v Carbopol Ultrez 10-NF in ibuprofen-glucosamine sulphate nanocrystals dispersion along with 0.01% methyl paraben and stirring solution overnight followed by pH adjustment from 5 to 6 using triethanolamine.

Experiment:
The laboratory scale-up of ibuprofen-glucosamine sulphate nanocrystals was carried out by dispersing ibuprofen (50-600 mg) in appropriate volume of methanol (5 mL) to form organic phase whereas, aqueous phase was prepared by dissolving glucosamine sulphate (5-10%) and poloxamer 407 in 20 mL distilled water. The organic phase was dropwise injected into aqueous phase maintained under continuous stirring with evaporation of methanol followed by probe sonication (Sonics Vibracell, USA) for 5 min with 10 sec on and 5 sec off cycle. The resultant nanocrystals were supplemented with 1% mannitol and stored at -80°C followed by lyophilization using freeze dryer (Alpha 1-2 L Basic, Martin Christ, Germany) to obtain powdered product. To evaluate the impact of ibuprofen on particle size of ibuprofen-glucosamine nanocrystals, batches were prepared with varying concentrations of ibuprofen and optimized (Table 1).
Table 1: Screening of amount of ibuprofen for the laboratory scale up of stable nanocrystals
Stabilizer
(1% or 100 mg) Amount of ibuprofen Stability Particle Size ±SD PDI±SD
Poloxamer 407 50 mg Stable 35.22±0.39 0.28±0.021
Poloxamer 407 100 mg Stable 34.57±0.79 nm 0.289±0.024
Poloxamer 407 200 mg Stable 108.24± 14.8 0.46±0.04
Poloxamer 407 600 mg Precipitate (Unstable) 1102.6±22 0.9±0.11

The ibuprofen-glucosamine sulphate nanocrystals prepared with 100 mg of ibuprofen was selected for further evaluation as it displayed monodispersed particles with size <100 nm, which is essential for improved skin permeation. The optimized ibuprofen-glucosamine sulphate nanocrystals displayed particle size of 34.57±0.79 nm with polydispersity index of 0.238±0.024, zeta potential of -2.81±0.6 mV and drug content of 7.05±0.19% with assay of 94±6.08% (Figure 2). The conversion of bulk ibuprofen into ibuprofen-glucosamine sulphate nanocrystals resulted into approximately ~90-fold increase in aqueous solubility (Figure 3). Apart from this, stability study performed at room temperature displayed significant (Two-way ANOVA, Dunnett's multiple comparison test) increase in particle size on 60th day due to crystallization followed by solubilization without any significant increase in PDI. Similarly, samples stored at refrigerated condition (2-8°C) demonstrated significant (Two-way ANOVA, Dunnett's multiple comparison test) increase in particle size after 14 days due to crystal growth followed by solubilization process with no significant change in PDI. These findings confirmed stability of ibuprofen-glucosamine sulphate nanocrystals for atleast 90 days stored at room temperature and refrigerated condition (Table 2).
Table 2: Stability study carried out for optimized ibuprofen-glucosamine sulphate nanocrystals at room temperature and refrigerated condition
Time (days) Room temperature Refrigerated temperature (2-8°C)
Average particle size (nm) PDI Average particle size (nm) PDI
0 32.70 ± 0.21 0.14 ±0.02 32.70 ± 0.21 0.14 ±0.02 ns
14 34.86 ± 0.53 ns 0.26 ± 0.009 ns 33.21±0.12 ns 0.186±0.064 ns
21 33.47 ± 0.28 ns 0.20 ± 0.031 ns 34.14±0.79 ns 0.218±0.022 ns
28 43.33 ± 1.02 **** 0.36 ± 0.004 ns 45.53±13.16 ** 0.235±0.009 ns
60 33.69 ± 5.7 ns 0.356 ± 0.1 ns 34.39± 5.75 ns 0.366± 0.01 ns
90 32.17 ± 2.3 ns 0.429 ± 0.023 ns 33.41±0.31 ns 0.161± 0.011 ns

Note: ns: P>0.05, **P<0.01, ****P<0.0001.
Following this, ibuprofen-glucosamine sulphate nanocrystals amalgamated topical gel was prepared by dispersing 2% w/v Carbopol Ultrez 10-NF along with 0.01% methyl paraben and allowed to stir overnight followed by pH adjustment from 5 to 6 using triethanolamine. The ibuprofen-glucosamine sulphate nanocrystals loaded gel exhibited drug loading of 0.507±0.029% with pH of 5.8±0.06. Apart from this, viscosity and spreadability of ibuprofen-glucosamine sulphate nanocrystals loaded topical gel were comparable with commercial diclofenac emulgel (Figure 4 and 5). Furthermore, ex-vivo permeation study was performed on rat skin displayed significantly high permeation for ibuprofen-glucosamine sulphate nanocrystals loaded gel (479.59±6.28 µg/cm2/h) as compared to ibuprofen gel (255.9±4.4 µg/cm2/h) (Figure 6).
In-vivo antiosteoarthritic potential for ibuprofen-glucosamine sulphate nanocrystals loaded gel was assessed using monosodium iodoacetate induced rat osteoarthritis model (Bhosale et al., 2024). The animal study was performed as per Committee for Control and Supervision of Experiments on Animals (CCSEA) guidelines, Ministry of Fishheries, Animal Husbandry, and Dairying, Government of India, New Delhi, India. The study was approved by the Institutional Animal Ethics Committee (IAEC) vide protocol #NIP/11/2022/PE/531. All the Wistar rats were acclimatized in animal house and divided into 7 groups each containing 6 animals (n=6). The details of the groups are as follow:
Groups Description Interventions
1 Normal control 50 µL of normal saline was injected by intra-articular route of administration
2 Positive control 3 mg/50 µL of monosodium iodoacetate through intraarticular route of administration to induce knee osteoarthritis
3 Treatment group 1 3 mg/50 µL of monosodium iodoacetate through intraarticular route + Conventional 5% w/w ibuprofen topical gel 2 times a day for 14 days
4 Treatment group 2 3 mg/50 µL of monosodium iodoacetate through intraarticular route + Conventional 10% glucosamine sulphate topical gel 2 times a day for 14 days
5 Treatment group 3 3 mg/50 µL of monosodium iodoacetate through intraarticular route + 0.5% w/w Ibuprofen nanocrystals loaded topical gel 2 times a day for 14 days
6 Treatment group 4 3 mg/50 µL of monosodium iodoacetate through intraarticular route + 0.5% w/w Ibuprofen and 10% w/w glucosamine sulphate nanocrystals loaded topical gel 2 times a day for 14 days
7 Treatment group 5 3 mg/50 µL of monosodium iodoacetate through intra-articular route + Blank Carbopol Ultrez 10-NF topical gel 2 times a day for 14 days

In brief, 3 mg/50 µL monosodium iodoacetate was injected into synovial cavity (right knee) of anaesthetized rats bent at right angle. After knee osteoarthritis induction, twice a day all the formulations were applied on shaved knee. Thickness of knee joint measured after monosodium iodoacetate injection displayed remarkable increment, thereby confirmed induction of osteoarthritis like conditions. Animals in all the treatment groups demonstrated a reduction in joint volume, out of which ibuprofen-glucosamine sulphate loaded gel displayed the most dramatic change (Figure 7).
At the end of 14th day, X-ray images were captured for right knee for exposure time of 10 s using Wipro GE-ELPRO operated at 100 mA and 50 kV. X-ray reports for all the animals were evaluated by following Kellgren-Lawrence scoring method.
Kellgren-Lawrence based scoring system evaluates the presence and severity of joint space narrowing and osteophytes. Where, Grade 0: No signs of osteoarthritis, Grade 1: Possible osteophytic lipping or doubtful narrowing of joint space, Grade 2: Definite osteophyte ad possible narrowing of joint space, Grade 3: Moderate multiple osteophytes, definite narrowing of joint space and possible deformity of bone ends and Grade 4: Large osteophytes, marked narrowing of joint space, severe sclerosis, and definite deformity of bone ends.
The result for X-ray analysis is illustrated in figure 8. Radiographic image of normal control displayed absence of joint space narrowing with no osteophyte formation. In contrast, rats injected with mono-sodium iodoacetate displayed joint space narrowing with osteophyte formation, thereby indicates the development of osteoarthritis like conditions. Rats treated with various other dosage forms displayed reduction in joint space narrowing and osteophyte formation. However, rats treated with ibuprofen-glucosamine sulphate nanocrystals loaded gel indicated absence of joint space narrowing and osteophytes.
Furthermore, histopathological evaluation for right knee was performed using toluidine blue and safranin-O/acid fast green staining. The Osteoarthritis Research Society International scoring for histology illustrated high score for positive control thereby indicating induction of osteoarthritis like conditions. Rats treated with ibuprofen gel and commercial diclofenac emulgel displayed reduction in knee joint inflammation as well as cartilage erosion. In contrast, rats treated with ibuprofen-glucosamine sulphate nanocrystals loaded gel demonstrated cartilage thickness recovery like normal control (Figure 9).
To evaluate the impact of treatment on the expression of inflammatory cytokines western blot analysis of synovial tissue was performed (Figure 10). The ratio of protein intensity to β-actin is expressed as fold change. The expression of COX-2, TNF-α and IL-1β in disease induced animals displayed respectively 9.01, 2.66 and 2.51-fold change as compared to normal control. Rats treated with conventional ibuprofen gel depicted 0.36, 0.57 and 0.83-fold reduction in COX-2, TNF-α and IL-1β respectively as compared to disease control. In comparison, ibuprofen nanocrystals loaded gel treatment resulted a 0.41, 1.38 and 1.16-fold reduction in COX-2, TNF-α and IL-1β, respectively as compared to disease control. As expected, animals treated with ibuprofen-glucosamine sulphate nanocrystals loaded gel demonstrated a 0.15, 0.24 and 0.35-fold reduction in COX-2, TNF-α and IL-1β levels, respectively. These findings advocate for improved anti-inflammatory as well as cartilage regenerating potential of ibuprofen-glucosamine sulphate nanocrystals loaded gel.
, Claims:We Claim:
1. A composition for management of joint pain and inflammation in osteoarthritis comprising:
a) ibuprofen present in amount of 50-600 mg;
b) methanol present in amount of 5 mL, wherein the ibuprofen in amount of 50-600 mg is mixed with the methanol in amount of 5 mL to form an organic phase;
c) glucosamine sulphate present in amount of 5-10%; and
d) a hydrophilic non-ionic surfactant, wherein an aqueous phase is prepared by dissolving the glucosamine sulphate present in amount of 5-10% in 20 mL of distilled water;
wherein the organic phase is dropwise injected into the aqueous phase under continuous stirring with evaporation of methanol followed by a probe sonication to form nanocrystals of the ibuprofen and the glucosamine sulphate.
2. The composition for management of joint pain and inflammation in osteoarthritis as claimed in claim 1, wherein the nanocrystals of the ibuprofen and the glucosamine sulphate is supplemented with a 1% mannitol and a stored at -80°C and followed by a lyophilization to obtain powdered form of the ibuprofen and glucosamine sulphate nanocrystals.
3. A method for synthesis of a composition for management of joint pain and inflammation in osteoarthritis comprising steps of:
a) dispersing ibuprofen in amount of 50-600 mg in methanol to form an organic phase;
b) dissolving glucosamine sulphate in amount of 5-10% with a hydrophilic non-ionic surfactant in 20 mL distilled water to form an aqueous phase;
c) injecting the organic phase obtained at step (a) dropwise injected into the aqueous phase obtained at step (b) under continuous stirring condition with evaporation of methanol;
d) agitating particles solution obtained at step (c) by a probe sonication for duration of a 5 min with a 10 sec on and a 5 sec off cycle to obtain nanocrystals; and
e) supplementing the nanocrystals obtained at step (d) with 1% mannitol and storing at temperature of -80°C followed by lyophilization to obtain powdered form of nanocrystals.
4. The method as claimed in claim 3, wherein amount of methanol is 5 mL for preparing an organic phase.
5. The method as claimed in claim 3, wherein the powdered form of nanocrystals of the ibuprofen and the glucosamine sulphate is synthesized into a gel form by steps of dispersing 2% w/v Carbopol Ultrez 10-NF along with 0.01% methyl paraben in ibuprofen-glucosamine sulphate nanocrystals dispersion and stirring overnight followed by pH adjustment from 5 to 6 using triethanolamine.

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