DESC:FORM 2
THE PATENTS ACT, 1970
(39 OF 1970)
&
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10; rule 13)

1. TITLE OF THE INVENTION – A NOVEL CAPSULE FORMULATION OF P-PROPOXY BENZOIC ACID
2. Applicant(s)
NAME: R K University
NATIONALITY: INDIAN
ADDRESS: R K University, School of Pharmacy, Bhavnagar Highway,
Tramba, Rajkot-360020, Gujrat, India

3. PREAMBLE TO THE DESCRIPTION

The following specification particularly describes the invention and the manner in which it is to be performed.

A NOVEL CAPSULE FORMULATION OF P-PROPOXY BENZOIC ACID
FIELD OF THE INVENTION
The present invention is all about a novel capsule formulation of p-Propoxy benzoic acid. The present invention also relates to a novel capsule formulation of p-Propoxy benzoic acid comprising p-Propoxy benzoic acid, binder, disintegrant and filler. The present invention also relates to a novel capsule formulation of p-Propoxy benzoic acid and process of preparing the same.

BACKGROUND OF THE INVENTION
Diabetes is a severe metabolic disorder which is indicated by hyperglycemia due to lack of insulin or the action of insulin on its target tissues or both. It is one of the major public health problems and is now becoming a global epidemic. In India, the prevalence of diabetes in the population is 8.9%. One in six people with diabetes in the world is from India. India is deemed as the world's capital of diabetes.

The synthetic oral anti-diabetic drugs and insulin currently used for the treatment of diabetes, are effective in controlling the elevated blood glucose levels but have various side effects and do not control the complications related to diabetes.

p- Propoxy benzoic acid is an organic compound that is also a part of several herbal anti-diabetic plants. It can have the potential of blocking certain targets such as a-amylase, a-glucosidase, and PTP1B. Inhibition of such enzymes can lead to anti-diabetic action. While discussing Type 2 Diabetes, a-amylase, PTP1B and a-glucosidase are emerging targets to provide synergistic effect against the disease.

p- Propoxy benzoic acid is found to give inhibitory effect on protein tyrosine phosphatases 1B (PTP1B) and a- glucosidase receptors which are responsible for inducing diabetes. Hence, it can be coined as multitarget antidiabetic drug.

Single-target oral hypoglycaemic agents target one specific enzyme responsible for glucose elevation and block it to produce hypoglycemia. Inhibition of a single target may result in less effective over time and fails to decrease glucose levels. Hence, the requirement for alternatives is mandatory. For that reason, multi-target oral hypoglycaemic agents can rectify this limitation and provide a potent anti-diabetic effect. From the results of the study, p-Propoxy benzoic acid (p-PBA) can be claimed as the first multi-target oral hypoglycaemic agent inhibiting a-amylase, a-glucosidase, and PTP1B enzyme. p-PBA was procured from a highly authentic source ensuring the purity and screening for anti-diabetic activity was conducted at various levels.

There is a need in the society to have heat stable and chemically stable composition of p-Propoxy benzoic acid which can be manufactured easily with less complexity, and having minimized side effects.

The inventors of the present invention surprisingly found a capsule formulation of p-Propoxy benzoic acid which is stable and present invention can overcome the above stated problems associated with the existing treatment.

OBJECTIVE OF THE INVENTION
The main objective of the present invention is a novel capsule formulation of p-Propoxy benzoic acid.

The other main objective of the present invention is a novel capsule formulation of p-Propoxy benzoic acid which is stable.

Another objective of the present invention is to provide a novel capsule formulation of p-Propoxy benzoic acid which is safe and having enhanced effectiveness.

Yet another objective of the present invention is to provide a novel capsule formulation of p-Propoxy benzoic acid which is having improved patient compliance.

Yet another objective of the present invention is to provide a novel capsule formulation of p-Propoxy benzoic acid which is useful in the treatment of diabetes.

SUMMARY OF THE INVENTION
The main aspect of the present invention is to provide a novel capsule formulation of p-Propoxy benzoic acid.

The other main aspect of the present invention is to provide a novel capsule formulation of p-Propoxy benzoic acid comprising p-Propoxy benzoic acid, binder, disintegrant and filler.

Another aspect of the present invention is to provide a novel capsule formulation of p-Propoxy benzoic acid and process of preparing the same.

The details of one or more aspect of the invention are set forth in the description below. Other features, objects and advantages of the invention will be apparent from the description.

DETAILED DESCRIPTION OF THE INVENTION
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described.

As used herein, the term "formulation" or “composition” unless otherwise defined refers to granules and/or solid oral pharmaceutical dosage forms or solid dispersions, suspension of the invention.

As used herein, whether in a transitional phase or in the body of a claim, the terms “comprise(s)” and “comprising” are to be interpreted as having an open-ended meaning. That is, the terms are to be interpreted synonymously with the phrases “having at least” or “including at least”. When used in the context of a process, the term “comprising” means that the process includes at least the recited steps, but may include additional steps. When used in the context of a composition, the term “comprising” means that the composition includes at least the recited features or components, but may also include additional features or components.

As used herein, the singular forms “a,” “an” and “the” specifically also encompass the plural forms of the terms to which they refer, unless the content clearly dictates otherwise.

The term “about” is used herein to means approximately, in the region of, roughly, or around. When the term “about” is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In general, the term “about” or “approximately” is used herein to modify a numerical value above and below the stated value by a variance of 20%.

The present invention overcomes the aforesaid drawbacks of the above, and other objects, features and advantages of the present invention will now be described in greater detail. Also, the following description includes various specific details and are to be regarded as merely exemplary. Accordingly, those of ordinary skill in the art will recognize that: without departing from the scope and spirit of the present disclosure and its various embodiments there may be any number of changes and modifications described herein.

The composition of present invention as described herein provide a novel capsule formulation.

As will be understood by one skilled in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible sub ranges and combinations of sub ranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, et cetera As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, et cetera As will also be understood by one skilled in the art all language such as “up to,” “at least,” and the like include the number recited and refer to ranges which can be subsequently broken down into sub ranges as discussed above.

As used in this document, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art.

The main embodiment of present invention is about a novel capsule formulation comprising p-Propoxy benzoic acid, binder, disintegrant and filler.

p-Propoxy benzoic acid:
p-Propoxy benzoic acid was procured from Thermo Fisher Scientific, Gloucester, United Kingdom (UK). p- Propoxy benzoic acid is an organic weak acid consisting carboxylic acid group on the para position. P- Propoxy benzoic acid was found to be the active chemical constituent of numerus herbal plants reported to give certain medicinal properties including anti-diabetic action. A report suggests the therapeutic activity of plant extract containing p-Propoxy benzoic acid against microbial growth. P-Propoxy benzoic acid is also reported to give aphrodisiac activity. Certain evidence on study performed on p-Propoxy benzoic acid suggests the utilization of this compound against cancer.

As per one embodiment the p-Propoxy benzoic acid is present in the composition in an amount from about 0.1 to 300 mg, preferably in the range from 1 to 300 mg, preferably in the range from 1 to 200 mg, more preferably in the range from 1 to 150 mg and most preferably in the range from 1 to 100 mg.

As per one embodiement, the binders are defined as a substance which is used to turn powder to granules; this is achieved through the process of granulation. During granulation, powder substances are accumulated to form larger particles called granules. Binders ensure that tablets, powders, granules and others can be formed with the required mechanical strength.

As per one embodiment, the binder is selected from PVP K30, HPMC E50 LV, HPMC K4M, (Poly ethylene glycol) PEG 6000, sodium alginate and Eudragit RL 100. In the present invention, most preferably binder is PVP-K30.

As per one embodiment of the present invention, the binder is present in the composition i in an amount from about 0.1 to 300 mg, preferably in the range from 1 to 300 mg, preferably in the range from 1 to 200 mg, more preferably in the range from 1 to 150 mg and most preferably in the range from 1 to 100 mg.

As per one embodiment, a disintegrant is defined as an additive that promotes disintegration, which is the breakage of a pharmaceutical formulation into small fragments when in contact with a liquid medium. In doing so, the surface area available for dissolution is increased and drug dissolution is accelerated.

As per one embodiment, the disintegrant is selected from chitosan hydrochloride, magnesium stearate, corn starch, calcium alginate, carboxymethyl cellulose and microcrystalline cellulose. In a most preferred embodiment, magnesium stearate is used as a disintegrant in the present invention.

As per one embodiment of the present invention, the disintegrant is present in the composition in an amount from about 0.1 to 200 mg, preferably in the range from 1 to 200 mg, preferably in the range from 1 to 150 mg, more preferably in the range from 1 to 100 mg and most preferably in the range from 1 to 50 mg.

As per one embodiment, fillers are defined as an excipients which are used to increase the volume of the material to enable easier processing of the ingredients and make it into a size suitable for consumption. Moreover, they can stabilize the product and help during manufacturing.

As per one embodiment, the filler is selected from talc, lactose, calcium salts and starch. In a most preferred embodiment, talc is used as a filler in the present invention.

As per one embodiment of the present invention, the filler is present in the composition in an amount from about 0.1 to 500 mg, preferably in the range from 1 to 500 mg, preferably in the range from 1 to 450 mg, more preferably in the range from 1 to 420 mg and most preferably in the range from 1 to 400 mg.

As per one embodiment, the present invention relates to a novel capsule formulation of p-Propoxy benzoic acid for oral administration.

As per one preferred embodiment, the process of preparing a novel capsule formulation of p-Propoxy benzoic acid comprising the steps of,
mixing p-Propoxy benzoic acid, talc and PVP-K30;
passing the wet mass of reaction mixture (a) through the number 14 mesh size sieve in order to produce wet granules;
drying the granules from step (b) in hot-air over at 50 ? for 2 hours;
mixing magnesium stearate with the granules for 10 minutes;
filling step (d) mixture in hard gelatin capsule.

As per one embodiment, p-Propoxybenzoic acid (p-PBA) is a first multi-target oral hypoglycaemic agent inhibiting a-amylase, a-glucosidase, and PTP1B enzyme. p-PBA used for anti-diabetic activity and was conducted at various levels.

The invention is further illustrated by the following examples which are provided to be exemplary of the invention and do not limit the scope of the invention. While the present invention has been described in terms of its specific embodiments, certain modifications and equivalents will be apparent to those skilled in the art and are intended to be included within the scope of the present invention.

EXAMPLES

EXAMPLE 1: A NOVEL CAPSULE FORMULATION OF P-PROPOXY BENZOIC ACID:
Ingredients Quantity

p-Propoxy benzoic acid 50 mg
PVP-K30 80 mg
Magnesium stearate 20 mg
Talc 250 mg
Table: 1 Formulation of a novel capsule formulation of p-Propoxy benzoic acid

Procedure:
p-Propoxy benzoic acid, talc and PVP-K30 were mixed;
the wet mass of reaction mixture (a) was passed through the number 14 mesh size sieve in order to produce wet granules;
the granules from step (b) were dried in hot-air over at 50 ? for 2 hours;
magnesium stearate was mixed with the granules for 10 minutes;
step (d) mixture was filled in hard gelatin capsule.

EXAMPLE 2: EVALUATION PARAMETERS OF HARD GELATIN CAPSULES CONTAINING P-PROPOXY BENZOIC ACID
PARAMETER Value (Mean ± SEM)
Bulk density (g/mL) 0.48 ± 0.07
Tapped density (g/mL) 0.587 ± 0.11
Hausner’s Quotient 1.15
Carr’s Index (%) 14.81
Angle of repose (°) 32.484 ± 8.21
Flow rate (g/s) 10.766 ± 1.62
Weight uniformity (mg) 398.727 ± 11.73
Content Uniformity (mg) 4.308 ± 0.697
Disintegration time 3.52 ± 0.81 min
Table: 2 Evaluation parameters of optimized formulation (as per example 1)

EXAMPLE 3: DISSOLUTION STUDY
Dissolution study was conducted using the paddle dissolution apparatus in which 0.1 N HCl solution was considered as the dissolution medium. The apparatus was filled with 900 mL of 0.1 N HCl which was maintained at 37.0 ± 1.0 ?. The capsule formulation (as per example 1) was incorporated attached with a small wire to prevent floating. 5 mL medium was taken out at every 5 minutes for the analysis and it was replaced with 5 mL of freshly prepared 0.1 N HCl. The samples were subjected to UV spectroscopy and absorbance was recorded at wavelength of 233 nm by using Shimadzu spectrophotometer.
The result of dissolution rate is given in below table:
Time (min) % Drug release
10 52.45
20 65.84
30 95.42
40 99.98
Table: 3 Dissolution rate results

EXAMPLE 4: a-AMYLASE INHIBITION
The study was carried out according to Johnson et al recommendations. 0.02 M sodium phosphate buffer was prepared and pH was adjusted to 7.4. 250 mg of soluble potato starch was added to 50 mL water and incubated for 15 minutes. 100 mL sodium phosphate buffer was added to 0.1 mg of a-amylase to prepare enzyme solution. Five different concentrations ranging from 20, 40, 60, 80 and 100 µg/mL solutions were prepared for acarbose (standard drug) and p-Propoxy benzoic acid (test drug). 1.0 g of 3, 5-dinitrosalicylic acid was added to 50 mL distilled water followed by drop wise addition of 30.0 g sodium potassium tartrate tetrahydrate, 20 mL of 2 N sodium hydroxide and diluted up to 100 mL distilled water to prepare 3,5-Dinitrosalicylic acid (DNS) reagent. For the conduction of assay, 1 mL of a-amylase was added to 1 mL of test/standard and incubated for 30 minutes. After the incubation period, 1 mL starch solution was added and furtherly incubated for 3 minutes. The reaction mixture was placed in closed tube and incubated at 85 ? for 15 minutes. The reaction mixture was cooled diluted with 9 mL distilled water and subjected to UV-spectroscopy for determination of absorbance at 540 nm. Percentage inhibition of a-amylase was calculated and recorded (Table 4) by the following equation:
Percentage inhibition=(Absorbance of control-Absorbance of test)/(Absorbance of control)*100

Concentration (µg/mL) Treatment groups
p-PBA Acarbose
20 35.48 44.27
40 42.03 59.95
60 55.38 71.86
80 62.90 78.32
100 70.25 87.10
Table: 4 a-amylase Percentage Inhibition

Half-Maximal inhibitory concentration (IC50) for p-PBA (24.82 µg/mL) and acarbose (56.59 µg/mL) was calculated as given in Table 5.
Concentration (µg/mL) IC50 Concentration (µg/mL)
Acarbose 24.82
p-PBA 56.59
Table: 5 Half-Maximal Inhibitory Concentration (IC50) against a-amylase

Half-maximal inhibitory concentration (IC50) calculated from the results of a-amylase inhibition assay p-PBA was 56.59 µg/mL and it also resulted in a significant percentage inhibition within the specific range of concentration compared to standard drug acarbose. Hence, the anti-diabetic activity of p-PBA has been confirmed in the in-vitro domain.

EXAMPLE 5: IN VIVO STUDY
36 healthy and adult Sprague-Dawley rats of either sex weighing 290 ± 20 g were kept for overnight fasting. Each animal was subjected to 290 mg/kg nicotinamide (0.9 % saline solution) intraperitonially prior to streptozotocin administration to reduce the mortality. After 15 minutes of nicotinamide administration, each animal was subjected to 65 mg/kg streptozotocin prepared in 0.1 M citrate buffer (4.15 pH) intraperitonially. After the administration of streptozotocin, animals were subjected to 10% glucose solution orally for 24 hours in order to reduce the mortality. Blood glucose level of animals were determined by retro orbital sinus and animals over 200 mg/kg glucose level were grouped accordingly for anti-diabetic study.

After inducing diabetic effect in rats, each healthy rats were distributed into six groups. Group I consisted of healthy rats treated with 0.1 % carboxymethyl cellulose served as normal control. Group II consisted of streptozotocin-nicotinamide type-II diabetic rats treated with 0.1% carboxy methyl cellulose and served as disease control. Group III was treated with 3 mg/kg glibenclamide and served as a standard group. Group IV was treated with p-PBA 100 mg/kg, Group V
was treated with p-PBA 200 mg/kg and Group VI was treated with p-PBA 300 mg/kg.
Group Name Treatment Received Dosage Route
I Normal control 0.1% Carboxy methyl cellulose 1 mg/kg Oral
II Disease Control (Diabetic) 0.1% Carboxy methyl cellulose 1 mg/kg Oral
III Glibenclamide (3 mg/kg) Glibenclamide 3 mg/kg Oral
IV p-PBA (100 mg/kg) p-Propoxy benzoic acid 100 mg/kg Oral
V p-PBA (200 mg/kg) p-Propoxy benzoic acid 200 mg/kg Oral
VI p-PBA (300 mg/kg) p-Propoxy benzoic acid 300 mg/kg Oral
Table: 6 Overview of study plan

Example 5.1
Effect of p-PBA formulation on Oral Glucose Tolerance Test (OGTT)
The beneficial effect of p-PBA in single-dose treatment was evaluated in a Phase-II study in which an oral glucose tolerance test (OGTT) was performed in healthy rats treated with different doses of p-PBA and 3 mg/kg glibenclamide. Results of OGTT in healthy rats shows significant decrease in blood glucose levels (mg/dL) overtime a period of 2 hours. The results show a statistically significant decrement with P value < 0.05. The results of the study suggest the glucose-lowering activity of p-PBA in healthy rats as shown in table 7.

Parameters Treatment groups
Normal Control Disease control Glibenclamide (3 mg/kg) p-PBA (100 mg/kg)b
p-PBA (200 mg/kg)
FPG (mg/dL) 112.5 ± 2.6 111.3 ± 6.4 114.3 ± 3.9 115.5 ± 3.1 115 ± 4.7
0.5-h PG (mg/dL) 109.5 ± 4.4 190.8 ± 6.7* 140.0 ± 9.1*# 165.25 ± 3.4*# 155.25 ± 6.5*#
1-h PG (mg/dL) 106.5 ± 3.7 180 ± 7.3* 137 ± 3.9*# 158.75 ± 3.4*# 144.75 ± 4.7*#
2-h PG (mg/dL) 107.5 ± 2.5 172.3 ± 7* 120 ±3.7*# 146.75 ± 2.7*# 137.50 ± 4.2*#
Table: 7 Oral glucose tolerance test in healthy rats

Data are represented as Mean ± SEM, p-PBA= p-Propoxy benzoic acid
FPG =Fasting Plasma Glucose
0.5-h PG = Plasma glucose after 0.5 hour of treatment
1-h PG = Plasma glucose after 1 hour of treatment
2-h PG = Plasma glucose after 2 hours of treatment
*Indicates statistically significant difference between normal control group and disease control group with P-Value <0.01
#Indicates statistically significant difference between Disease control group and treatment groups with P-value <0.01

Example 5.2
Effect of p-PBA formulation on body weight
The body weight of each group of animals was measured by an animal weighing scale (Kents scientific) from Week 0 to Week 4. The average weight of animals of each group was recorded as Mean ± SEM. The average reduction in body weight of Type-II diabetic rats was maximum in disease control group as compared to other treated groups as shown in table 8.
Group Body Weight
Week 0 Week 1 Week 2 Week 3 Week 4
Normal Control 265.5 ± 9.07 266.5 ± 9.50 273.5 ± 6.51 286.2 ± 10.48 287.5 ± 9.97
Disease Control 245.5 ± 6.12 243.7 ± 6.25 237.2 ± 5.51* 219.5 ± 6.08*# 210.0 ± 4.7*#
Glibenclamide (3 mg/kg) 254.5 ± 7.96 253.5 ± 7.05 246.25 ± 5.41 246.0 ± 6.78*# 244.0 ± 4.24*#
p-PBA (100 mg/kg)b
264.2 ± 9.63 257.0 ± 9.68 251.0 ± 8.28 245.0 ± 9.21*# 233.0 ± 6.15*#
p-PBA (200 mg/kg) 240.0 ± 5.58 238.25 ± 3.71 232.75 ± 3.99* 225.7 ± 3.07* 216.5 ± 2.1*
p-PBA (300 mg/kg) 249.0 ± 6.56 246.2 ± 6.25 238.0 ± 6.77* 233.2 ± 7.71*# 227.2 ± 7.5*#
Table: 8 Trend of Body Weight over Treatment Period

*Indicates statistically significant difference in treatment groups in comparison of normal control group with P-value < 0.05.
#indicates statistically significant difference in treatment groups in comparison of disease control group with P-value < 0.05.
Example 5.3
Effect of p-PBA formulation on blood glucose levels
Plasma glucose levels remained similar over time period of 28 days in normal control group. The statistically significant difference was observed in comparison between disease control and normal control group with P-value < 0.001. Plasma glucose levels remained elevated in all groups on Day 0. From Day 7 to Day 28, the levels of plasma glucose levels started to decrease significantly in test treatment groups compared to disease control group with P-value < 0.001. Plasma glucose levels collected from Day 0 to Day 28 has been recorded as given in Table 9.
Treatment group Blood glucose levels (mg/dL)
Day 0 Day 7 Day 14 Day 28
Normal Control 114.7 ± 4.21 114.2 ± 2.17 112 ± 4.88 117.7 ± 4.87
Disease Control 213.7 ± 4.42* 227.5 ± 4.05* 247.7 ± 4.53* 255.5 ± 3.59*
Glibenclamide (3 mg/kg) 217.5 ± 5.19* 189.7 ± 4.55*# 167.2 ± 3.45*# 147.2 ± 4.27*#
p-PBA (100 mg/kg) 222.2 ± 3.35* 202.2 ± 4.01*# 182.7 ± 2.43*# 175.5 ± 2.96*#
p-PBA (200 mg/kg) 222.0 ± 4.18* 200.2 ± 2.93*# 189.7 ± 2.63*# 166.5 ± 3.30*#
p-PBA (300 mg/kg) 217.75 ± 4.57* 191.0 ± 5.52*# 172.7 ± 2.59*# 152.0 ± 4.24*#
Table: 9 Reduction in Blood Glucose Levels Over the Treatment Period

*Indicates statistically significant difference in treatment group in comparison of normal control group with p-value < 0.001.
#Indicates statistically significant difference in treatment group in comparison of disease control group with p-value < 0.001.

Example 5.4
Effect of p-PBA formulation on glycosylated haemoglobin and serum insulin levels
Measured levels of Glycosylated haemoglobin (HbA1c) increased significantly in disease control group compared to normal control group with P-value < 0.001. Treatment of Type-II diabetic rats with p-PBA groups significantly regulated the HbA1c levels. The levels of HbA1c in treatment groups were significantly reduced compared to disease control group with P-value < 0.001. The statistical significance difference (P-value < 0.05) was observed in p-PBA (200 mg/kg) and p-PBA (300 mg/kg) in comparison with p-PBA (100 mg/kg).

Levels of serum insulin significantly reduced in disease control group in comparison to normal control group with P-value < 0.001. Reduction in serum insulin levels were regulated by p-PBA (300 mg/kg), p-PBA (200 mg/kg) and p-PBA (100 mg/kg) significantly with P-value < 0.001. The levels of HbA1c and serum insulin is given for different groups in Table 10.
Group HbA1c (mg/g of Hb%) Serum Insulin (mcU/mL)

Normal Control 0.39 ± 0.02 20.94 ± 0.46
Disease Control 1.81 ± 0.17$ 4.69 ± 0.42$
Glibenclamide (3 mg/kg) 0.51 ± 0.02*# 18.83 ± 0.21*
p-PBA (100 mg/kg) 1.13 ± 0.06* 12.75 ± 0.43*
p-PBA (200 mg/kg) 0.73 ± 0.04*# 15.45 ± 0.32*
p-PBA (300 mg/kg) 0.59 ± 0.03*# 18.12 ± 0.33*
Table: 10 Levels of Glycosylated Haemoglobin and Serum Insulin

HbA1c = Glycosylated Haemoglobin
*Indicates statistically significant difference between disease control group and treatment groups with p-value < 0.001.
#Indicates statistically significant difference between p-PBA (100 mg/kg) and other treatment groups with p-value <0.05.
$Indicates statistically significant difference between normal control group and disease control group with p-value <0.001.

Example 5.5
Effect of p-PBA formulation on Blood glucose level at different time interval
The effect of p-PBA on diabetic rats was evaluated which resulted in a significant decrement in blood glucose level over the time period of 24 hours as compared to disease control group.

Blood glucose level Treatment groups
Normal Control Disease Control Glibenclamide (3 mg/kg) p-PBA (100 mg/kg) p-PBA (200 mg/kg) p-PBA (300 mg/kg)
0 hours (mg/dL) 120 ± 4.9 213.2 ± 3.5* 210 ± 4.4* 221.4 ± 5.4* 218 ± 6.6* 215.4 ± 4.9*
0.5 hours (mg/dL) 115 ± 5.2 225.6 ± 8.1* 196.2 ± 4.1* 209 ± 2.9* 201.5 ± 5.9* 200.4 ± 3.8*
2 hours (mg/dL) 107.4 ± 4.4 223 ± 6.0* 175.6 ± 4.7*# 197.9 ± 4.1*# 189.3 ± 3.7*# 183.5 ± 4.4*#
3 hours (mg/dL) 106 ± 2.5 215.6 ± 4.9* 162.9 ± 6.2*# 187.4 ± 3.9*# 176.4 ± 5.0*# 169.2 ± 3.2*#
6 hours
(mg/dL) 114.9 ± 4.0 219 ± 3.8* 145.3 ± 5.4*# 173.1 ± 4.0**# 163.6 ± 4.7*# 157 ± 5.1*#
12 hours (mg/dL) 112.7 ± 3.9 215.9 ± 4.2* 160 ± 3.8*# 184.9 ± 3.6*# 170.8 ± 8.2*# 166.4 ± 3.5*#
24 hours
(mg/dL) 110 ± 6.5 215.3 ± 5.6* 174.4 ± 5.8*# 201.4 ± 5.3*# 188 ± 6.4*# 182.3 ± 5.0*#
Table: 11 Blood glucose level at different time interval

*Indicates statistically significant difference between normal control group and treatment groups with P-value < 0.001
**Indicates statistically significant difference between disease control group and treatment groups with P-value < 0.005

EXAMPLE 6: IN-SILICO STUDY
Example 6.1: Molecular docking studies
On the initial basis, inhibitory activity of p-propoxybenzoic acid against computational model of a-amylase, a-glucosidase and Protein tyrosine phosphatase 1B (PTP1B) was measured by obtaining binding energy of interaction from blind docking. Purpose of these studies was to minimize unnecessary discomfort to experimental animals.
Molecular docking studies were performed by utilizing Auto Dock Tools. Input PDB file for p-propoxybenzoic acid was prepared. Target compound was docked against the a-amylase enzyme, a-glucosidase enzyme and PTP1B. In each docking, the receptor protein was kept rigid and protein polar hydrogens were added. A total of 20 blind docking runs were carried out against each enzyme. The output files were extracted and evaluated for the binding energy of the enzyme-ligand interaction. p-PBA reported significant correlation in binding affinity towards a amylase, a-glucosidase and PTP1B in in-silico molecular docking study. Results of docking studies are collected and recorded as given in Table 12.
Enzyme Ligand Binding energy of individual docking runs (kcal/mol) Docking score (kcal/mol)
1 2 3 4 5 6 7 8 9 10
a-amylase Acarbose (standard) 7.92 8.47 8.95 8.89 9.04 8.98 8.80 8.76 8.44 8.67 8.69 ± 0.34
p-PBA 6.93 7.45 7.32 6.94 7.85 7.98 7.73 8.20 7.92 7.82 8.43 ± 0.44*
a- glucosidase Acarbose (standard) 8.90 9.91 9.56 9.45 9.04 9.42 9.93 8.96 9.70 9.31 9.42 ± 0.37
p-PBA 7.43 8.34 8.50 7.98 8.33 8.10 8.24 9.15 9.03 8.35 9.19 ± 0.49#
PTP1B Standard Drug X 9.95 10.25 10.11 10.29 9.70 9.95 9.89 10.25 10.11 10.08 10.05 ± 0.19
p-PBA 8.14 8.02 8.24 8.18 9.01 8.66 8.92 8.95 9.21 7.94 9.40 ± 0.47$
*Indicates the statistical correlation of test drug with Acarbose with P-value <0.0001
#Indicates the statistical correlation of test drug with Acarbose with P-value <0.0001
$Indicates the statistical correlation of test drug with standard X with P-value <0.0001
PTP1B = Protein tyrosine phosphatase 1B
Standard drug X = (3-({5-[(n-acetyl-3-{4-[(carboxycarbonyl)(2-carboxyphenyl) amino]-1-naphthyl}-l-alanyl)amino]pentyl}oxy)-2-naphthoic acid) ,CLAIMS:CLAIMS:
We claim,
1. A novel capsule formulation of p-Propoxy benzoic acid comprising p-Propoxy benzoic acid, binder, disintegrant and filler.
2. The novel capsule formulation of p-Propoxy benzoic acid as claimed in claim 1, wherein the binder is selected from PVP K30, HPMC E50 LV, HPMC K4M, (Poly ethylene glycol) PEG 6000, sodium alginate and Eudragit RL 100.
3. The novel capsule formulation of p-Propoxy benzoic acid as claimed in claim 1, wherein the disintegrant is selected from chitosan hydrochloride, magnesium stearate, corn starch, calcium alginate, carboxymethyl cellulose and microcrystalline cellulose.
4. The novel capsule formulation of p-Propoxy benzoic acid as claimed in claim 1, wherein the filler is selected from talc, lactose, calcium salts and starch.
5. The novel capsule formulation of p-Propoxy benzoic acid as claimed in claim 1, wherein the p-Propoxy benzoic acid used is in the range from 1 to 100 mg.
6. The novel capsule formulation of p-Propoxy benzoic acid as claimed in claim 1, wherein the binder used is in the range from 1 to 100 mg.
7. The novel capsule formulation of p-Propoxy benzoic acid as claimed in claim 1, wherein the disintegrant used is in the range from 1 to 50 mg.
8. The novel capsule formulation of p-Propoxy benzoic acid as claimed in claim 1, wherein the filler used is in the range from 1 to 400 mg.
9. The novel capsule formulation of p-Propoxy benzoic acid as claimed in claim 1, wherein the process of preparation comprises the steps of,
a) mixing p-Propoxy benzoic acid, talc and PVP-K30;
b) passing the wet mass of reaction mixture (a) through the number 14 mesh size sieve in order to produce wet granules;
c) drying the granules from step (b) in hot-air over at 50 ? for 2 hours;
d) mixing magnesium stearate with the granules for 10 minutes;
e) filling step (d) mixture in hard gelatin capsule.

Dated this 06th Jul, 2022