Description:
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 TABLET FORMULATION OF SALICIN
2. Applicant(s)
NAME: RK UNIVERSITY
NATIONALITY: INDIAN
ADDRESS: RK University, Bhavnagar Highway, Kasturbadham, Rajkot-360020, Gujarat, 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 TABLET FORMULATION OF SALICIN
FIELD OF THE INVENTION
The present invention is related to a novel tablet formulation of salicin. The present invention particularly relates to a novel tablet formulation of salicin for treating asthma.

BACKGROUND OF THE INVENTION
Asthma is a major non-communicable disease (NCD), affecting both children and adults. Inflammation and narrowing of the small airways in the lungs cause asthma symptoms, which can be any combination of cough, wheeze, shortness of breath and chest tightness.

Asthma affected an estimated 262 million people in 2019 and caused 461000 deaths. Asthma is often under-diagnosed and under-treated, particularly in low- and middle-income countries. People with under-treated asthma can suffer sleep disturbance, tiredness during the day, and poor concentration.

The structural changes of the airways associated with asthma, broadly referred to as airway remodeling, is a pathological feature of chronic asthma that contributes to the clinical manifestations of the disease. Airway remodeling in asthma constitutes cellular and extracellular matrix changes in the large and small airways, epithelial cell apoptosis, airway smooth muscle cell proliferation, and ?broblast activation.

Angiogenesis is the formation of new blood vessels as an extension of pre-existing vessels. Under conditions of homeostasis, a balance exists between angiogenic activators and inhibitors, and a state of vascular quiescence is maintained in which there is no net change in vascularization.

Patients with asthma are no longer maintaining vascular quiescence in the bronchial wall and thus have reached a pro-angiogenic state. This pathological angiogenesis occurs due to overproduction of angiogenic factors, underproduction of inhibitors, or a combination of each of these issues, leading to increased vascularization. Increased numbers of blood vessels in the bronchial wall is strongly correlated to the severity of asthma. Increased vascularity in the airway and the increased vessel permeability which occurs concurrently contribute to the thickening of the inner airway wall and the development of airway edema. These symptoms lead to narrowing of the airway lumen which reduces airflow and leads to the obstructive symptoms of asthma.

Vascular endothelial growth factor (VEGF) is a signaling protein and the main regulator of endothelial cell growth. It is released by many cells that stimulate the formation of blood vessels and is a classic pro fibrotic growth factor. The overexpression of VEGF in the lung can lead to significant airway remodeling. Therefore, antagonizing VEGF is important in the prevention of remodeling and in preserving epithelial barrier function. VEGF has significant roles in inflammation, angiogenesis and the vascular permeability of airways and in subepithelial collagen deposition, airway smooth muscle hyperplasia and the development of physiological abnormalities of the airway.

Salicin is an alcoholic ß-glucoside. It is a biosynthetic precursor to salicyl aldehyde. It is closely related chemically to aspirin. It has two part glycone (beta -D-glycosyl) and aglycone (salicyl alcohol) which is responsible for anti-inflammatory activity. Major source of salicin is willow (salix) bark. Willow bark has been used for many years for anti-inflammatory, antipyretic, and analgesic effects. Efficacy of willow bark is believed to be due to salicin content.

Salicin blocks angiogenesis by blocking VEGF pathway. This anti-angiogenic activity can be helpful in asthma as inhibition of angiogenic factors inhibits airways remodeling ultimately leading to anti-inflammatory effects in asthma. Salicin works by reducing the production of pro-angiogenic factors, prevent them binding to their receptors or block their actions. Salicin inhibits reactive oxygen species production and the extracellular signal-regulated kinase pathway Reactive oxygen species play a critical role in angiogenesis and regulate intracellular signaling pathways.

Rachid Berair et al in their article, Asthma Therapy and Its Effect on Airway Remodelling, discloses that the detrimental pathophysiological consequences of some features of remodeling, like increased airway smooth muscle mass and subepithelial fibrosis, are well documented. However, whether targeting these by therapy would be beneficial is unknown. Bronchial thermoplasty is the only asthma therapy to primarily target remodelling. To date, no asthma drug therapy has been used to primarily target airway remodelling.

Hence there is need to develop formulation which can directly targets airway remodelling. The inventors of the present invention have surprisingly formulated a tablet of salicin which gives anti angiogenic effect in asthmatic patients which directly targets airway remodelling.

OBJECTIVE OF THE INVENTION
The main objective of the present invention is to provide a novel tablet formulation of salicin.

Another objective of the present invention is to provide a novel tablet formulation of salicin for treating asthma.

Yet another object of the present invention is to provide a novel tablet formulation of salicin which has better patient compliance.

One other object of the present invention is to provide a novel tablet formulation of salicin which is targeted formulation which gives direct effect to airway remodeling in asthmatic patients.

SUMMARY OF THE INVENTION
The main aspect of the present invention is to provide a novel tablet formulation of salicin.

Another main embodiment of the present invention is to provide a novel tablet formulation of salicin comprising salicin, diluent, binder, disintegrant, lubricant and anticaking agent.

The other aspect of the present invention is to provide a novel tablet formulation of salicin and process for preparing the same.

DESCRIPTION OF THE INVENTION
The main embodiment of the present invention is to provide a novel tablet formulation of salicin.

The detailed description set forth below is intended as a description of exemplary embodiments and is not intended to represent the only forms in which the exemplary embodiments may be constructed and/or utilized. The description sets forth the functions and the sequence of steps for constructing and/or operating the exemplary embodiments. However, it is to be understood that the same or equivalent functions and sequences which may be accomplished by different exemplary methods are also intended to be encompassed within the spirit and scope of the invention.

As defined herein, all scientific and technical 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 process 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 stated in the present invention 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.

As stated herein, that it follows in a transitional phrase 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.

The main embodiment of the present invention is to provide a novel tablet formulation of salicin.

Another main embodiment of the present invention is to provide a novel tablet formulation of salicin comprising salicin, diluent, binder, disintegrant, lubricant and anticaking agent.

The salicin was procured online from selleckchem.com, USA.
As per another embodiment, the salicin is used in the present invention is preferably in the range of 100 to 500mg, more preferably of 100 to 300 mg and most preferably in the range of 100-200 mg.

As per one embodiment, diluent used in the present invention can be selected from powdered cellulose, microcrystalline cellulose, silicified microcrystalline cellulose, lactose, starch, dibasic calcium phosphate, tribasic calcium phosphate, calcium carbonate, dextrates, dextrin, dextrose, kaolin, magnesium carbonate, magnesium oxide, sugars such as sucrose; sugar alcohols such as mannitol, sorbitol, erythritol. As per preferred embodiment, microcrystalline cellulose is used as diluent.

As per one embodiment, diluent used is preferably in the range of 30-50 mg, more preferably of 35-50 mg and most preferably of 40-50 mg.

As per one embodiment, binder used in the present invention can be selected from hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, carbomers, carboxymethylcellulose sodium, dextrin, ethyl cellulose, methylcellulose, shellac, zein, gelatin, polymethacrylates, polyvinyl pyrrolidone (PVP K30), pregelatinized starch, sodium alginate. As per preferred embodiment, pregelatinized starch is used as binder.

As per one embodiment, diluent used is preferably in the range of 20-40 mg, more preferably of 25-40 mg and most preferably of 30-40 mg.

As per one embodiment, disintegrant used in the present invention can be selected from carboxymethylcellulose and its sodium salt, crospovidone, starch, sodium starch glycolate, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, anhydrous colloidal silica, low-substituted hydroxypropyl cellulose and sodium alginate. As per preferred embodiment, anhydrous colloidal silica is used as disintegrant.

As per one embodiment, disintegrant used is preferably in the range of 5-10 mg, more preferably of 5-8 mg and most preferably of 5-7 mg.

As per one embodiment, lubricant used in the present invention can be selected from magnesium stearate, calcium stearate, zinc stearate; stearic acid, hydrogenated vegetable oil, hydrogenated castor oil, glyceryl palmitostearate, glyceryl behenate, polyethylene glycols, corn starch, sodium stearyl fumarate, sodium benzoate, mineral oil, talc. As per preferred embodiment, stearic acid is used as lubricant.

As per one embodiment, lubricant used is preferably in the range of 1-6 mg, more preferably of 1-5 mg and most preferably of 2-5 mg.

As per one embodiment, anticaking agent used in the present invention can be selected from D-mannitol, colloidal silicon dioxide, calcium silicate, magnesium silicate, and calcium stearate, talc. As per preferred embodiment, talc is used as anticaking agent.

As per one embodiment, anticaking agent used is preferably in the range of 5-15 mg, more preferably of 5-12 mg and most preferably of 7-12 mg.

As per one embodiment, the process of preparing a novel tablet formulation of salicin comprises the step of:
a) Weighing salicin and diluent;
b) Passing salicin and diluent of step (a) through the sieve and mixing;
c) Blending of mixture of (b) with blender;
d) Weighing of other ingredients;
e) Passing ingredients of step (d) through the sieve and mixing;
f) Blending mixture of step (d) with mixture of step (c) with blender;
g) Compressing the mixture of step (f) into tablets;
h) Packing of the final tablet formulation.
As per one other embodiment, the present invention provides a novel tablet formulation of salicin for treating asthmatic patients by giving direct effect on airway remodeling.

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 TABLET FORMULATION OF SALICIN
Sr. No. Ingredients Quantity (in mg)
1 Salicin 200
2 Microcrystalline Cellulose 45
3 Pregelatinized starch 35
4 Anhydrous colloidal silica 06
5 Stearic acid 04
6 Talc 10
7 Water q.s
Table 1: Formulation of a novel tablet of salicin
Process of preparation:
a) Weighing salicin and microcrystalline cellulose;
b) Passing salicin and microcrystalline cellulose of step (a) through the sieve and mixing;
c) Blending of mixture of (b) with blender;
d) Weighing of other ingredients;
e) Passing ingredients of step (d) through the sieve and mixing;
f) Blending mixture of step (d) with mixture of step (c) with blender;
g) Compressing the mixture of step (f) into tablets;
h) Packing of the final tablet formulation.

EXAMPLE 2: EVALUATION OF PHYSICAL PARAMETERS OF A NOVEL TABLET FORMULATION OF SALICIN
2.1: Weight variation of tablets
20 tablets were randomly taken from each batch and weighed using the analytical weighing balance. The weight of each tablet and the average weight of tablet was calculated. The weight of individual tablet was compared with the average weight of tablet. The results were considered as positive as not more than two tablets fell outside the limits.

2.2: Hardness of tablets
The hardness of the tablet was evaluated by a Monsanto tablet hardness tester. The hardness was evaluated for 20 tablets and hardness values were recorded.

2.3: Friability of tablets
The friability of the tablet was evaluated using Roche Friabilator. The test was conducted for twenty tablets and placed in friabilator chamber. The chamber was allowed to rotate 100 revolutions. The friability values were recorded.

2.4: Diameter and Thickness of tablets
Diameter and thickness of tablet was measured using Vernier caliper and the test was replicated for 20 tablets.

Results:
Sr. No. Parameter Mean + SEM P-value
1 Weight Uniformity (mg) 301 ± 0.3 0.008
2 Hardness (kg/cm2) 7.45 ± 0.02 0.01
3 Friability (%) 0.19 -
4 Diameter (mm) 12.42 ± 0.09 0.04
5 Thickness (mm) 1.91 ± 0.05 0.005
Table 2: Physical parameters of salicin tablets

EXAMPLE 3: DRUG RELEASE STUDY OF A NOVEL TABLET FORMULATION OF SALICIN
Dissolution study on the formulation was performed as per the IP procedure in calibrated dissolution test apparatus equipped with paddles employing 900 mL of 0.1 N HCl for initial 2 hours and pH 7.4 PBS for the remaining period of time as a medium. The paddles were rotated at 50 rpm and temperature was maintained at 37 ± 1 ?. 5 mL of samples were withdrawn at regular time intervals for 12 hours and replaced with same volume of freshly prepared dissolution medium to maintain the constant volume of medium throughout the experiment. The samples were diluted with PBS and drug content was determined by measuring the absorbance at 280 nm using UV spectrometer. Cumulative amount of drug released was plotted against the time period to calculate the rate of drug release.

Result:
Sr. No. Time (Hrs) Cumulative % drug released
1 1 9.87
2 2 21.45
3 3 34.82
4 4 48.51
5 5 62.48
6 6 81.11
7 7 90.24
8 8 99.87
Table 3: % CDR of Salicin from Tablets
Salicin has substantial drug release in dissolution medium over 8 hours of time period due to the solubility of salicin in wide range of mediums including 0.1 N HCl and phosphate buffer. 50% of drug release was obtained in approximately 4 hours.

EXAMPLE 4: IN-VITRO STUDY (ANTI-HAEMOLYTIC STUDY)
Anti-haemolytic assay of salicin was performed as described previously in Yang et al, 2005 with modification.
Initially, 10 mL of blood was withdrawn from Wistar Albino rat by retro orbital puncher and collected in EDTA vials. The vial was centrifuged for 5 minutes as 1000 x g. The supernatant was removed and the solution was washed three times with 0.2 M phosphate buffer solution to prepare erythrocyte suspension. 1 mL Erythrocyte suspension was transferred into the Eppendorf. Total 6 Eppendorf were prepared containing 1 mL suspension in each. Additionally, 0.5 % saline solution was prepared and added to the sample tubes. 0.5 mL of five salicin solutions having concentrations of 200 µg/mL, 400 µg/mL, 600 µg/mL, 800 µg/mL and 1000 µg/mL prepared in phosphate buffer solution were added to 5 tubes containing erythrocyte suspension.
The samples were incubated at room temperature for 20 minutes. Further, 0.5 mL of hydrogen peroxide solution was added into each tube to provoke haemolytic activity. The samples were centrifuged at 1000 x g for 10 minutes. After 10 minutes, the absorbance of each sample was measured at 265nm. The absorbance was also measured for the tube containing only erythrocyte suspension considered as the negative control. The percentage inhibition in haemolysis was calculated with the reference of negative control. Quercetin (200-1000 µg/mL) was taken as the standard drug to compare the anti-haemolytic activity of salicin. The IC50 value for both drugs were calculated from the linear equation.
Result:
Sr No. Concentration
(µg/mL) % Inhibition
Salicin Quercetin
1 200 52.70 39.19
2 400 60.14 43.92
3 600 77.70 60.14
4 800 85.81 65.54
5 1000 97.97 86.49
Table 4: Anti-haemolytic activity of Salicin and Quercetin

Sr No. Compound IC50 Concentration (µg/mL)
1 Quercetin 172.12
2 Salicin 444.25
Table 5: Half-maximal inhibitory concentration (IC50) against Quercetin

Hydrogen peroxide solution induced the haemolysis in erythrocyte suspension. The extent of haemolysis was maximum in negative control and the percent inhibition of haemolysis decreased as the concentration of salicin increased. The trend was compared with the standard drug quercetin. The calculated IC50 value of test drug salicin justify the anti-haemolytic activity.

EXAMPLE 5: IN-VIVO STUDY
Experimental Animals
The Institute Animal Ethics Committee (IAEC) authorized the use of 36 adult Wistar Albino rats of either sex weighing 240 ± 15 g, which were acquired from the Animal House of the School of Pharmacy, R.K. University. Prior to the studies, they were acclimatized. Animals were given regular food and water ad libitum and were kept in standard ambient settings.

5.1: Acute in-vivo screening model: Haloperidol Induced Catalepsy
Tests for catalepsy measures that how long a mouse can remain in a fixed posture with its forelimbs fully extended and resting on a 4-centimeter-high bar (1.0 cm diameter). When both front paws are withdrawn from the bar or the animal moves its head in an inquisitive way, catalepsy is deemed to be over. For every additional 20 seconds that the enforced position is held for, the severity of the cataleptic behavior is increased by one point.
A standard bar test was used to measure catalepsy in albino rats (n=6) after inducing it with haloperidol (1.0 mg/kg, i.p.) for 30 minutes and then again after 120 minutes and 240 minutes.

30 Wister stain of rats will be divided into 5 groups of 6 animals. Each group was treated with respective compound.
Group I: 0.5 % CMC solution (10 mL/kg) (p.o.)
Group II: 0.5% CMC solution (10 mL/kg) (p.o.)
Group III: Trihexyphenidyl (10 mg/kg) (p.o.)
Group IV: Salicin (100 mg/kg) (p.o.) (L)
Group V: Salicin (200 mg/kg) (p.o.) (M)
Group VI: Salicin (300 mg/kg) (p.o) (H)
After 30 minutes 1 mg/kg haloperidol will be given to Group II to VI rats intra-peritoneally.
The time required (latency) for the animal to remove its forelimbs from an elevated horizontal bar was measured.

Result:
Sr. No. Group Cataleptic score
30 Min 60 Min 120 Min 180 Min
1 Normal Control 0 ± 0 0 ± 0 0 ± 0 0 ± 0
2 Negative control 57.75 ± 4.35 62.5 ± 3.70 54.25 ± 4.79 48.25 ± 2.99
3 Standard 31.5 ± 3.42* 30.00 ± 3.56* 34.25 ± 3.3* 34.00 ± 5.6**
4 Salicin (L) 49 ± 2.94*# 46.75 ± 2.22*# 51.5 ± 2.65*# 46.75 ± 2.99**#
5 Salicin (M) 43.75 ±2.22*# 41 ± 2.16*# 46.00 ± 5.48*# 44.25 ± 3.30**#
6 Salicin (H) 39.75 ± 2.22*# 37.75 ± 4.99*# 40.75 ± 2.63*$ 41.75 ± 2.75**$
Table 6: Difference in cataleptic scores of treatment group over time
Data is represented as mean ± SEM
*Indicates statistical significance of treatments compared to negative control with P-value < 0.001
**Indicates statistical significance of treatments compared to negative control with P-Value < 0.01
#Indicates statistical significance of salicin groups compared to standard with P-value < 0.001
$indicates statistical significance of salicin groups compared to standard with P-value < 0.05

Cataleptic score of normal control group was zero which is understandable as in normal condition rat cannot stand still on the glass rod. As the animals were treated with haloperidol, the catalepsy was observed and it was maximum in negative control group without any subsequent treatment. Cataleptic score was minimum in standard group having comparable results with treatment groups. As the dosage of salicin increased the cataleptic score decreases which justify the beneficial activity against catalepsy induced by haloperidol.

5.2: Sub-acute in-vivo screening model: Histamine Induced Bronchospasm
Initially, the animals were divided into 5 groups of 6 animals in each as below.
Group I: 1 mL Distilled water (p.o.)
Group II: Chlorpheniramine maleate (2 mg/kg) (p.o.) + Histamine (0.2%)
Group III: Salicin (100 mg/kg) (p.o.) + Histamine (0.2%)
Group IV: Salicin (200 mg/kg) (p.o.) + Histamine (0.2%)
Group V: Salicin (300 mg/kg) (p.o.) + Histamine (0.2%)

On day 0, all groups were treated with aerosol of 0.2% histamine aerosol and the baseline pre-convulsion time (s) was measured for each group before the administration of treatment. All groups were treated with respective drugs for 14 days and again subjected to 0.2% histamine aerosol to measure pre-convulsion time.

Result:
The difference and percentage difference of pre-convulsion time (PCT) was calculated and presented in table below.
Sr. No. Group Pre-convulsion time Difference % difference
Day 0 Day 14
1 Normal control 110 ± 3.56 111.5 ± 2.65 3 ± 3.56 2.64 ± 3.09
2 Chlorpheniramine Maleate (2mg/kg) 102.25 ± 2.06 211.25 ± 8.77* 109 ± 10.8 51.50 ± 3.02
3 Salicin (100 mg/kg) 109 ± 2.94 144.75 ± 10*# 35.75 ± 7.63 24.51 ± 3.81
4 Salicin (200 mg/kg) 104.25 ± 4.57 165.75 ± 7*# 61.5 ± 5.07 37.08 ± 2.20
5 Salicin (300 mg/kg) 107.25 ± 3.95 186.25 ± 5.32*# 79 ± 3.16 42.42 ± 1.24
Table 7: Variation in PCT time over time period of 14 days
Data is represented as mean ± SEM
*Indicates statistical significance between treatment groups and untreated group (p-value < 0.001)
#Indicates statistical significance between standard treatment group and test group (p-value <0.001)

Initially the animals were familiarized with histamine aerosol. The values of pre-convulsion time observed on day 0 were similar to each other. After the treatment period of 14 days with respective treatments, the pre-convulsion time decreased significantly in treatment group. The percentage difference between the pre-convulsion time of day 0 and day 14 was notable and maximum in standard drug Chlorpheniramine maleate followed by higher dose of salicin, medium dose of salicin and lower dose of salicin.

5.3: Chronic in-vivo screening model: Ovalbumin Induced Lung inflammation

Induction of lung inflammation
36 healthy Wistar Albino rats were divided into 6 groups of 6 in each. The experimental protocol was followed according to Dong et al. The animals were divided as per below.
Group I: 1 mL Distilled water (p.o.)
Group II: 1 mL Distilled water (p.o.) + 40 mg/kg Ovalbumin + Al(OH)2 2 mg/kg
Group III: Chlorpheniramine maleate (2 mg/kg) (p.o.) + 40 mg/kg Ovalbumin + Al(OH)2 2 mg/kg
Group IV: Salicin (100 mg/kg) (p.o.) + 40 mg/kg Ovalbumin + Al(OH)2 2 mg/kg
Group V: Salicin (300 mg/kg) (p.o.) + 40 mg/kg Ovalbumin + Al(OH)2 2 mg/kg

In order to induce lung inflammation each group expect group I were treated with 40 mg/kg ovalbumin and 2 mg/kg aluminum hydroxide intraperitoneally on 0th day, 7th day and 14th day to induce lung inflammation.

For next 14 days, the animals were treated with respective treatment and by end the end of the treatment period they were euthanized and dissected for histopathological study and blood parameter retrieval. Blood parameters including TNF-a, absolute WBC count, absolute eosinophil count and absolute neutrophil count were recorded.

Result:
Sr. No. Group TNF-a (pg/mL)
1 Normal Control 46.88 ± 5.48
2 Negative Control 715.1 ± 23.36#
3 Standard 117.78 ± 12.57*
4 Salicin (100 mg/kg) 577.25 ± 27.77*
5 Salicin (200 mg/kg) 237.95 ± 12.85*
6 Salicin (300 mg/kg) 152.48 ± 7.10*
Table 8: TNF-a levels of various groups
Data are represented as mean ± SD
*Indicates statistical significance of treatments compared to negative control with P-value < 0.001
#Indicates statistical significance of disease control compared to Normal control with P-value < 0.001

Sr. No. Group Absolute WBC count Absolute Eosinophil count Absolute Neutrophil count
1 Normal Control 7.4075 ± 0.22 0.4050 ± 0.15 0.1350 ± 0.05
2 Disease Control 18.9275 ± 2.38* 0.7050 ± 0.11* 0.3150 ± 0.06*
3 Standard 11.1125 ± 1.02*# 0.4000 ± 0.08# 0.1600 ± 0.03*
4 Salicin (L) 16.0300 ± 0.85* 0.6126 ± 0.07 0.2400 ± 0.02!*
5 Salicin (M) 14.6775 ± 0.57*# 0.5475 ± 0.03 0.2000 ± 0.02^#
6 Salicin (H) 12.86 ± 0.34*# 0.4450 ± 0.04 0.1700 ± 0.01#
Table 9: WBC, AEC, ANC levels of various groups
Data are represented as mean ± SD
*Indicates statistical significance of treatments compared to normal control with P-value < 0.01
#Indicates statistical significance of treatments compared to disease control with P-value < 0.01
^indicates statistical significance of treatment compared to normal control with P-value < 0.05
!indicates statistical significance of treatment compared to the disease control with P-value < 0.05

The sole aim of this screening model was to determine the extent of reduction in lung inflammation in presence of salicin. TNF-a, which is noticeable factor of inflammation was greatly increased in negative control group and reduced significantly in salicin treatment groups. Similarly, the absolute counts of WBC, eosinophil and Neutrophil were increased in disease control group and significantly reduced in treatment groups which clearly justifies the anti-inflammatory activity of salicin against asthma.

After performing, in-vitro and in-vivo study, it can be concluded that novel tablet formulation of salicin gives satisfactory results which proves that salicin tablet has antiasthmatic potential.
, C , Claims:CLAIMS
We claim;
1) A novel tablet formulation of salicin comprising 100 to 500 mg of salicin, 30-50 mg of diluent, 20-40 mg of binder, 5-10 mg of disintegrant, 1-6 mg of lubricant and 5-15 mg of anticaking agent.
2) The novel tablet formulation of salicin as claimed in claim 1 wherein said diluent is selected from powdered cellulose, microcrystalline cellulose, silicified microcrystalline cellulose, lactose, starch, dibasic calcium phosphate, tribasic calcium phosphate, calcium carbonate, dextrates, dextrin, dextrose, kaolin, magnesium carbonate, magnesium oxide, sugars such as sucrose; sugar alcohols such as mannitol, sorbitol, erythritol.
3) The novel tablet formulation of salicin as claimed in claim 1 wherein said binder is selected from hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, carbomers, carboxymethylcellulose sodium, dextrin, ethyl cellulose, methylcellulose, shellac, zein, gelatin, polymethacrylates, polyvinyl pyrrolidone (PVP K30), pregelatinized starch, sodium alginate.
4) The novel tablet formulation of salicin as claimed in claim 1 wherein said disintegrant is selected from carboxymethylcellulose and its sodium salt, crospovidone, starch, sodium starch glycolate, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, anhydrous colloidal silica, low-substituted hydroxypropyl cellulose and sodium alginate.
5) The novel tablet formulation of salicin as claimed in claim 1 wherein said lubricant is selected from magnesium stearate, calcium stearate, zinc stearate; stearic acid, hydrogenated vegetable oil, hydrogenated castor oil, glyceryl palmitostearate, glyceryl behenate, polyethylene glycols, corn starch, sodium stearyl fumarate, sodium benzoate, mineral oil, talc.
6) The novel tablet formulation of salicin as claimed in claim 1 wherein said anticaking agent is selected from D-mannitol, colloidal silicon dioxide, calcium silicate, magnesium silicate, and calcium stearate, talc.
7) The novel tablet formulation of salicin as claimed in claim 1 wherein said formulation of salicin is prepared by direct compression.
8) The novel tablet formulation of salicin as claimed in claim 1 wherein said tablet formulation comprising 200 mg salicin, 45 mg diluent, 35 mg binder, 6 mg disintegrant, 4 mg lubricant and 10 mg anticaking agent.
9) A process of preparation of novel tablet formulation of salicin as claimed in claim 1 comprises the step of:
a) Weighing salicin and diluent;
b) Passing salicin and diluent of step (a) through the sieve and mixing;
c) Blending of mixture of (b) with blender;
d) Weighing of other ingredients;
e) Passing ingredients of step (d) through the sieve and mixing;
f) Blending mixture of step (d) with mixture of step (c) with blender;
g) Compressing the mixture of step (f) into tablets;
h) Packing of the final tablet formulation.