Description:Preamble to the Description
[0001] The following specification describes the invention and the manner in which it is to be performed:
DESCRIPTION OF THE INVENTION
Technical field of the invention
[0001] The present invention relates to a composition of neem leaf water-soluble bitters to be used as a natural active in dermo cosmetic and nutraceutical applications. The invention also discloses a process of preparation of neem leaf water-soluble bitters which yields a composition of 100% aqueous solubility. The composition is cost effective, safe and non-toxic.
Background of the invention
[0002] Azadirachta indica commonly known as neem, is a natural herb recognized for several therapeutic properties. Neem has been traditionally used for several ailments for thousands of years. Neem modulates numerous biological processes without the occurrence of any adverse effects.
[0003] Neem comprises abundant phytochemical constituents including isoprenoids and non-isoprenoids. The bark, wood, sap, leaves, flowers, fruits, seeds, and oil of the neem tree are used for varied medicinal purposes.
[0004] The properties demonstrated by the constituents of neem include immunomodulatory, anti-inflammatory, antihyperglycaemic, antiulcer, antimalarial, antifungal, antibacterial, antiviral, antioxidant, antimutagenic and anticarcinogenic properties.
[0005] Neem has been used as a raw material in several applications of the dermo cosmetic industries. The extracts of neem have been incorporated into face wash, scrub, shampoo, oils, soaps and so on. Neem aids in removal of skin impurities and revitalizes the skin.
[0006] The water-soluble bitters have been profoundly used in cosmetic industries. The solubility of the bitters in water is limited. The neem leaf bitters are soluble in various organic solvents including alcohol and ether. The water insoluble bitters have reduced shelf life and are thermolabile.
[0007] The Patent Application US201916436267A entitled “Process for Preparing Water Soluble Azadiractin Powder Formulation with In-Built and External Inclusion Complexes” discloses a process for producing water soluble azadirachtin from neem seed kernel. The neem seed kernel comprises azadirachtin, limonoids and polysaccharide-protein matrix. The formulation relates to a free-flowing powder and is not chemically bound. The inclusion complex of the formulation comprises a derivative of cyclodextrin and a polysaccharide-protein mixture. The concentration of azadirachtin in the formulation is found to be 600 ppm. The process for the production of water-soluble azadirachtin comprises steps of removal of husk from the seeds, de-oiling of the seeds, extraction at neutral pH and liquid phase separation. The process further involves concentration of clarified aqueous extract, addition of external azadirachtin, filtration and spray drying to obtain water-soluble azadirachtin powder.
[0008] The Patent Application BR102017014693A entitled “Process for obtaining dry, water-soluble extract, azadriachtin a concentrate, from seeds of the species azadirachta indica (neem)” discloses process for extraction and purification of azadirachtins from Azadirachta indica seeds. The process yields water-soluble azadirachtins at a higher concentration. The process comprises steps of pre-treatment of the raw material to achieve the size range of mesh 4 to mesh 140, extraction with alcohols and esters, solvent removal, homogenization and phase separation. The solvents utilized for the extraction are ethanol and ethyl acetate. The process further comprises steps of concentration of azadirachtin to obtain dry, water-soluble powder of the extract. The process is compatible with green chemistry principles and does not incorporate any harmful solvents. The process results in consumption of less energy.
[0009] The Patent Application EP20060005864 entitled “Cosmetic composition comprising botanical extracts and a thickener” discloses composition in the form of emulsion in aqueous phase comprising a cross-linked polymer and a hydrophilic botanical extract. The composition comprises water-soluble anionic acrylic monomers, water-soluble non-ionic acrylate monomers, bifunctional monomeric cross-linking agent. The composition further comprises hydrophilic botanical extracts of seabuckthom, rose hip fruit, ginko biloba leaf, panax ginseng root and neem leaf. The concentration of hydrophilic botanical extract is in the range of 0.0001% to 1%. The composition comprises hydrophilic vitamin components including vitamin B3, provitamin B5 and vitamin C. The composition is used as a cosmetic for regulating the condition of skin.
[0010] Although neem has been incorporated in diverse applications, the solubility of the extracts in water is limited. Water being a universal solvent, the extracts that are soluble in water can be prepared instantly. Also, the water-soluble extracts have increased shelf life and are thermostable. Hence, there is a necessity for a process for the preparation of water-soluble neem extract with increased solubility.
Summary of the invention
[0011] The present invention overcomes the drawbacks of the existing neem leaf bitter compositions. The process for preparation yields higher concentrations of neem leaf water-soluble bitters. The neem leaf water-soluble bitters exhibit 100% aqueous solubility.
[0012] The present invention discloses a composition of neem leaf water-soluble bitters and a process of preparation thereof. The process of preparation of neem leaf water-soluble bitters comprises steps of mixing polyvinylpyrrolidone K 30 at a concentration of 27.5 % w/w and polyoxyethylene sorbitan monooleate at a concentration of 2.5% w/w in 500 ml of ethanol, adding 70% neem leaf hydroalcoholic extract to the mixture, evaporating the alcohol from the mixture, filtering the mixture to remove insoluble material and spray drying at 120 °C -140 °C to obtain greenish brown neem leaf water-soluble bitters powder.
[0013] The present invention further discloses a composition of neem leaf water-soluble bitters. The composition comprises neem leaf hydroalcoholic extract at a concentration of 70% w/w, polyvinyl pyrrolidone K-30 (PVP K-30) at a concentration of 27.5% w/w and polyoxyethylene sorbitan monooleate at a concentration of 2.5% w/w.
[0014] The neem leaf water-soluble bitter composition exhibits 100% aqueous solubility. The enhanced solubility is achieved by the addition of PVP K-30, a solubilizing agent and polyoxyethylene sorbitan monooleate, a surfactant. The neem leaf water-soluble bitter composition of the present invention comprises flavonoids at a concentration range between 8% w/v and 15% w/v.
[0015] The neem leaf water-soluble bitter composition of the present invention exhibits antioxidant activity at a minimal concentration. The neem leaf water-soluble bitter composition exhibits glutathione reductase activity, which aids in the pro-oxidant activity in chemoprevention of cancer cells. The neem leaf water-soluble bitter composition further exhibits hypoglycemic effect by inhibiting the activity of α-amylase and α-glucosidase enzymes.
[0016] The neem leaf water-soluble bitter composition of the present invention comprising bioactive components including flavonoids exhibit anti-inflammatory activity by the inhibition of cyclooxygenase-2 (COX-2) enzyme. Further, the neem leaf water-soluble bitter composition inhibits the growth of acne causing bacteria Propionibacterium acnes and Staphylococcus epidermidis. The neem leaf water-soluble bitter composition exhibits anti-cancer activity by the inhibition of proliferation of cancer cell lines.
[0017] The present invention discloses a composition of neem leaf water-soluble bitters and a process of preparation thereof. The process of preparation yields in 100% aqueous soluble neem leaf water-soluble bitter composition. The neem leaf water- soluble bitter composition is safe and non-toxic.

Brief description of the drawings
[0018] The foregoing and other features of embodiments will become more apparent from the following detailed description of embodiments when read in conjunction with the accompanying drawings. In the drawings, like reference numerals refer to like elements.
[0019] FIG 1 illustrates the flowchart for the process of preparing neem leaf water-soluble bitters.
[0020] FIG 2 tabulates the composition of neem leaf water-soluble bitters.
[0021] FIG 3 illustrates the percentage of radical scavenging activity of the neem leaf water-soluble bitter composition.
[0022] FIG 4 illustrates the percentage glutathione reduction by the neem leaf water-soluble bitter composition.
[0023] FIG 5 illustrates percentage α-amylase inhibition by the neem leaf water-soluble bitter composition.
[0024] FIG 6 illustrates percentage α-glucosidase inhibition by the neem leaf water-soluble bitter composition.
[0025] FIG 7 illustrates the percentage COX-2 inhibition of the neem leaf water-soluble bitter composition.
[0026] FIG 8 illustrates the zone of inhibition of the acne bacteria against neem leaf water-soluble bitter composition.
[0027] FIG 9 tabulates the minimum inhibitory concentration of the neem leaf water-soluble bitter composition.
[0028] FIG 10 illustrates the percentage inhibition of cell proliferation of MDA-MB-468 cell line by the neem leaf water-soluble bitter composition.
[0029] FIG 11 illustrates the percentage inhibition of cell proliferation of MDA-MB-468 cell line by the neem leaf water insoluble bitters.
[0030] FIG 12 illustrates the percentage inhibition of cell proliferation of MDA-MB-231 cell line by the neem leaf water-soluble bitter composition.
[0031] FIG 13 illustrates the percentage inhibition of cell proliferation of MDA-MB-231 cell line by the neem leaf water insoluble bitters.
[0032] FIG 14 illustrates the percentage inhibition of cell proliferation of Hek293T cell line by the neem leaf water-soluble bitter composition.
[0033] FIG 15 illustrates the percentage inhibition of cell proliferation of Hek293T cell line by the neem leaf water insoluble bitters.
[0034] FIG 16 illustrates the percentage inhibition of cell proliferation of HFF-1 cell line by the neem leaf water-soluble bitter composition.
[0035] FIG 17 illustrates the percentage inhibition of cell proliferation of HFF-1 cell line by the neem leaf water insoluble bitters.
Detailed description of the invention
[0036] In order to more clearly and concisely describe and point out the subject matter of the claimed invention, the following definitions are provided for specific terms, which are used in the following written description.
[0037] The term “Anti-inflammatory” refers to a substance which reduces inflammation in the body.
[0038] The term “Surfactant” refers to a substance which reduces the surface tension of a liquid.
[0039] The term “Water-soluble bitters” refers to the component of the botanical extract soluble in water.
[0040] The present invention discloses a composition of neem leaf water-soluble bitters and a process of preparation thereof. The composition comprises neem leaf hydroalcoholic extract, polyvinyl pyrrolidone K-30 and polyoxyethylene sorbitan monooleate. The process of preparation yields in a composition exhibiting enhanced water solubility.
[0041] FIG 1 illustrates the flowchart for the process of preparing neem leaf water-soluble bitters. The process (100) of preparing neem leaf water-soluble bitters begins with step (101) of mixing polyvinyl pyrrolidone K-30 (PVP K-30) at a concentration of 27.5% w/w and polyoxyethylene sorbitan monooleate at a concentration of 2.5% w/w. The mixture is stirred continuously in ethanol at room temperature until a clear solution is obtained (102). At step (103), neem leaf hydroalcoholic extract is added at a concentration of 70% w/w to the PVP K-30 and polyoxyethylene sorbitan monooleate ethanol mixture. The ethanol mixture is stirred continuously to obtain a clear solution. At step (104), the ethanol mixture is subjected to evaporation by a rotary evaporator. The evaporated mixture is added with 500 ml of distilled water to get a clear solution. At step (105), the evaporated mixture is subjected to filtration to remove insoluble material from the mixture. At step (106), the filtered mixture is subjected to spray drying at 120°C -140°C using spray dryer to obtain solubilized neem leaf water-soluble bitters. The neem leaf water-soluble bitters exhibit 100% aqueous solubility. The concentration of total flavonoids in the neem leaf water-soluble bitter composition is in the range of 8% w/v -15% w/v.
[0042] FIG 2 tabulates the composition of neem leaf water-soluble bitters. The composition comprises 70% of neem leaf hydroalcoholic extract, 27.5% of PVP K-30 and 2.5% of polyoxyethylene sorbitan monooleate. The composition exhibits 100% aqueous solubility in water. The enhanced solubility is achieved by the addition of PVP K-30, a solubilizing agent and polyoxyethylene sorbitan monooleate a surfactant. The neem leaf water-soluble bitter composition completely solubilizes in water. The aqueous solubility enhances the dissolution of the neem leaf water-soluble composition hence increasing the absorption in cosmetic applications. Additionally, the neem leaf water-soluble bitter composition is thermostable.
[0043] Having generally described this invention, a further understanding can be obtained by reference to certain specific examples, which are provided herein for purposes of illustration only and are not intended to be limiting unless otherwise specified.
Example 1: Analysis of antioxidant activity of the neem leaf water-soluble bitter composition
[0044] The antioxidant activity is performed to analyze the free radical scavenging activity of the composition. The flavonoids and water-soluble bitters of the composition aid in minimizing the effect of reactive oxygen species (ROS), free radicals, deoxyribose nucleic acid (DNA) damage and ageing.
[0045] The antioxidant activity was screened by the analysis of conversion of free radical 1,1-diphenyl-2-picryl hydroxyl (DPPH) to 1,1-diphenyl-2-picryl hydrazine. DPPH is a purple-colored substance and upon the action of antioxidants DPPH gets converted into colorless 1,1-diphenyl-2-picryl hydrazine. The change of color was measured by the rate of absorbance at 517 nm. The antioxidant activity was comparatively analyzed for neem leaf water-soluble bitters and neem leaf water insoluble bitters at concentrations of 1 µg/µl, 2.5 µg/µl, 5 µg/µl and 10 µg/µl. The control for the analysis was ascorbic acid at concentrations of 200 µg/µl, 400 µg/µl, 600 µg/µl, 800 µg/µl and 1000 µg/µl. The addition of DPPH to neem leaf water-soluble bitters, neem leaf water-insoluble bitters and ascorbic acid was followed by incubation for 15 minutes at room temperature. The absorbance was measured, and respective percentage of inhibition was calculated.
[0046] FIG 3 illustrates the percentage of radical scavenging activity of the neem leaf water-soluble bitter composition. The neem leaf water-soluble bitter exhibits maximum percentage of radical scavenging activity. The IC50 of neem leaf water-soluble bitters was found to be ~42.34 µg/ml and the IC50 for neem leaf water-insoluble bitters was found to be ~98.43 µg/ml. The neem leaf water-soluble bitters exhibit antioxidant activity at a minimal concentration of ~42.34 µg/ml.
Example 2: Analysis of glutathione reduction activity of the neem leaf water-soluble bitter composition
[0047] The glutathione reductase activity was performed to analyze the pro-oxidant activity of the composition. The generation of free radicals enables ROS-induced cell damage in cancerous cells. The analysis of reduced glutathione (GSH) and oxidized glutathione (GSS) relates to the generation of free radicals.
[0048] The generation of free radicals was measured by the reaction of Elman’s reagent with oxidized glutathione at 412 nm. The concentrations of neem leaf water-soluble bitters and neem leaf water insoluble bitters used were 1 µg/µl and the concentration of ascorbic acid standard used was 1mg/ml.
[0049] FIG 4 illustrates the percentage glutathione reduction of the neem leaf water-soluble bitter composition. The standard ascorbic acid indicates ~7% glutathione reduction. The neem leaf water insoluble bitters indicate ~20% glutathione reduction. The neem leaf water-soluble bitters indicate ~33% glutathione reduction. The increased percentage glutathione reduction relates to the pro-oxidant activity in chemoprevention of cancer cells.
Example 3: Analysis of α-amylase and α-glucosidase inhibitory activity of the neem leaf water-soluble bitter composition
[0050] The inhibitory activity of α-amylase and α-glucosidase were analyzed for the hypoglycemic effect of the neem leaf water-soluble bitter composition. The flavonoids present in the composition aid in management of diabetes mellitus.
[0051] The activity of α-amylase was analyzed by the addition of starch azure as a substrate to the neem leaf water-soluble bitters at a concentration of 1 mg/ml. The absorbance was measured at 540 nm. The percentage of inhibition was calculated by the measurement of absorbance.
[0052] FIG 5 illustrates percentage α-amylase inhibition by the neem leaf water-soluble bitter composition. The neem leaf water insoluble bitters exhibit percentage α-amylase inhibition at a concentration of 8 µg/ml whereas the neem leaf water-soluble bitters exhibit percentage α-amylase inhibition at a concentration of 28 µg/ml indicating the hypoglycemic effect of the composition.
[0053] FIG 6 illustrates percentage α-glucosidase inhibition by the neem leaf water-soluble bitter composition. The neem leaf water insoluble bitters exhibit percentage α-glucosidase inhibition at a concentration of 14 µg/ml whereas the neem leaf water-soluble bitters exhibit percentage α-glucosidase inhibition at a concentration of 42 µg/ml indicating the hypoglycemic effect of the composition.
Example 4: Analysis of anti-inflammatory activity of the neem leaf water-soluble bitter composition
[0054] The anti-inflammatory activity of the neem leaf water-soluble bitter composition was analyzed for the production of inflammatory mediators by the bioactive components of neem. The onset of inflammation results in activation of cyclooxygenase pathway where arachidonic acid gets converted to prostaglandins. The reaction is mediated by cyclooxygenase-2 (COX-2) enzyme. The screening of COX-2 enzyme correlates to the anti-inflammatory activity of the neem leaf water-soluble bitter composition. The analysis was performed by COX-2 inhibitor screening kit for neem leaf water-insoluble bitters and neem leaf water-soluble bitters.
[0055] FIG 7 illustrates the percentage COX-2 inhibition by the neem leaf water-soluble bitter composition. Neem leaf water insoluble bitters exhibited 8.9% inhibition of COX-2 enzyme. Neem leaf water-soluble bitters exhibited 47% inhibition of COX-2 enzyme. The presence of bioactive components including flavonoids of the neem leaf water-soluble bitter composition exhibits anti-inflammatory activity by the inhibition of COX-2 enzyme.
Example 5: Analysis of anti-acne activity of the neem leaf water-soluble bitter composition
[0056] The anti-acne activity of the neem leaf water-soluble bitter composition was evaluated to analyze the effect of neem leaf water-soluble bitter composition on the growth of acne-causing bacteria. Acne is commonly caused by the growth of Propionibacterium acnes and Staphylococcus epidermidis. The prevalence of acne has been increased due to the increase in antibiotic resistance. The analysis of anti-acne activity was evaluated by agar well diffusion assay.
[0057] The agar well diffusion assay for Propionibacterium acnes was performed on brain heart infusion (BHI) medium. The inoculums of the bacteria were prepared and incubated for 48 hours on BHI agar. The agar gels were cut by sterile cork borer and the neem leaf water-soluble bitter composition was added to the wells. The agar gels were incubated anaerobically for 72 hours at 37o C. The zone of inhibition was evaluated and hence minimum inhibitory concentration was calculated.
[0058] The agar well diffusion assay for Staphylococcus epidermidis was performed on nutrient agar medium. The suspension cuture of the bacteria was prepared with 0.5ml of the bacterial suspension in the molten agar. The agar gels were cut by sterile cork borer and the neem leaf water-soluble bitter composition was added to the wells. The agar gels were incubated anaerobically for 72 hours at 37o C. The zone of inhibition was evaluated and hence minimum inhibitory concentration was calculated.
[0059] FIG 8 illustrates the zone of inhibition of the acne bacteria against neem leaf water-soluble bitter composition. FIG 9 tabulates the minimum inhibitory concentration of the neem leaf water-soluble bitter composition. The agar well diffusion assay indicates the antibacterial activity of the neem leaf compoisition against Propionibacterium acnes and Staphylococcus epidermidis. The neem leaf water-soluble bitter composition exhibited minimum inhibitory concentration of 13.2 ± 0.2 mm against Propionibacterium acnes. The neem leaf water-soluble bitter composition exhibited minimum inhibitory concentration of 11.9 ± 0.2 mm against Staphylococcus epidermidis.
[0060] Example 6: Analysis of in-vitro anticancer activity of neem leaf water-soluble bitter composition
[0061] The in-vitro anti-cancer activity was evaluated to analyze the effect of neem leaf water-soluble bitter composition on proliferation of cancerous cells. The evaluation also analyzes the effectiveness of the neem leaf water-soluble bitter composition on improving the immune response, eliminating free radicals, and inhibiting cell division and inflammation.
[0062] The in-vitro anti-cancer activity of the neem leaf water-soluble bitter composition was evaluated by 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. The MTT assay determines the mitochondrial activity of living cells. The MTT assay was performed on breast cancer cell lines MDA-MD-231 and MDA-MB-468. The concentrations of neem leaf water insoluble bitters and neem leaf water-soluble bitters used were 0.1 mg/ml, 0.25 mg/ml, 0.5 mg/ml, and 1 mg/ml. Dimethyl sulphoxide (DMSO) was used as the negative control and doxorubicin was used as the positive control for the assay. The cells were cultured for 48 and 72 hours and the absorbance was measured at 570 nm.
[0063] FIG 10 illustrates the percentage inhibition of cell proliferation of MDA-MB-468 cell line by the neem leaf water-soluble bitters. FIG 11 illustrates the percentage inhibition of cell proliferation of MDA-MB-468 cell line by the neem leaf water insoluble bitters. The corresponding IC50 value for neem leaf water insoluble bitters was found to be ~0.4 mg/ml. The corresponding IC50 value for neem leaf water-soluble bitters was found to be ~0.09 mg/ml. The neem leaf water-soluble bitters exhibits higher percentage of inhibition of cell proliferation at a lower concentration.
[0064] FIG 12 illustrates the percentage inhibition of cell proliferation of MDA-MB-231 cell line by the neem leaf water-soluble bitters. FIG 13 illustrates the percentage inhibition of cell proliferation of MDA-MB-231 cell line by the neem leaf water insoluble bitters. The corresponding IC50 value for neem leaf water insoluble bitters was found to be ~0.38 mg/ml. The corresponding IC50 value for neem leaf water-soluble bitters was found to be ~0.55 mg/ml. The neem leaf water-soluble bitters exhibits higher percentage of inhibition of cell proliferation at a lower concentration.
Example 7: Analysis of safety evaluation of the neem leaf water-soluble bitter composition
[0065] The evaluation of safety dosage was determined to analyze the therapeutic usage of the neem leaf water-soluble bitter composition. The neem leaf water-soluble bitter composition was evaluated for the safety dosage by in-vitro toxicity studies against Kidney Epithelial cells (Hek293T) and Skin Fibroblast cells (HFF-1). The cytotoxicity displayed determines the cell growth, function and survival.
[0066] The safety analysis was performed by MTT assay which indicates the mitochondrial activity of living cells. The Hek293T and human foreskin fibroblast (HFF-1) cells were incubated overnight. The incubated cells were added with the neem leaf water-insoluble bitters and neem leaf water-soluble bitters at concentrations of 0.1 mg/ml, 0.25 mg/ml, 0.5 mg/ml, and 1 mg/ml. DMSO was used as the negative control and doxorubicin was used as the positive control for the assay. The cells were cultured for 48 and 72 hours and the absorbance was measured at 570 nm.
[0067] FIG 14 illustrates the percentage inhibition of cell proliferation of Hek293T cell line by the neem leaf water-soluble bitters. FIG 15 illustrates the percentage inhibition of cell proliferation of Hek293T cell line by the neem leaf water insoluble bitters. The neem water insoluble bitters were found to be safe and non-toxic against Hek293T cells with an IC50 value of ~ 0.85 mg/ml. The neem water insoluble bitters were found to be safe and non-toxic against Hek293T cells with an IC50 value of ~ 1.95 mg/ml.
[0068] FIG 16 illustrates the percentage inhibition of cell proliferation of HfF-1 cell line by the neem leaf water-soluble bitters. FIG 17 illustrates the percentage inhibition of cell proliferation of HFF-1 cell line by the neem leaf water insoluble bitters. The neem water insoluble bitters were found to be safe and non-toxic against HF-1 cells with an IC50 value of ~ 6.8 mg/ml. The neem water insoluble bitters were found to be safe and non-toxic against HFF-1 cells with an IC50 value of ~ 7.2 mg/ml.
[0069] The present invention discloses a composition of neem leaf water-soluble bitters and a process of preparation thereof. The composition comprising hydroalcoholic neem extract, PVP K-30 and polyoxyethylene sorbitan monooleate exhibits enhanced water solubility. The enhanced water solubility aids in feasibility of preparing an extract along with prolonged shelf-life and thermostability. The aqueous solubility enhances the dissolution of the neem leaf water-soluble composition hence increasing the absorption in cosmetic applications. The neem leaf water-soluble bitter composition further comprises crucial bioactive components including flavonoids at a higher concentration. The neem leaf water-soluble bitter composition exhibits essential therapeutic properties including antioxidant, anti-acne, anti-cancer, anti-inflammatory, and anti-hyperglycemic properties.
, Claims:We Claim:

1. A process for the preparation of neem leaf water-soluble bitters, the process (100) comprising steps of:
a) mixing 27.5% of the solubilizing agent and 2.5% of the surfactant in 500 ml of ethanol at room temperature (101);
b) subjecting the mixture to continuous stirring until a clear solution is obtained (102);
c) adding 70% of the neem leaf hydroalcoholic extract to the mixture with stirring (103);
d) subjecting the mixture to evaporation by an evaporator (104);
e) subjecting the evaporated mixture to filtration to remove insoluble material (105); and
f) spray drying the mixture to obtain solubilized neem leaf water-soluble bitters (106).

2. The process (100) as claimed in claim 1, wherein said solubilizing agent is polyvinylpyrrolidone K 30 (PVP K-30).

3. The process (100) as claimed in claim 1, wherein said surfactant used is polyoxyethylene sorbitan monooleate.

4. The process (100) as claimed in claim 1, wherein said spray drying is achieved by a spray dryer at a temperature range of 120°C -140°C.

5. The process (100) as claimed in claim 1, wherein said product is greenish-brown neem leaf water-soluble bitters exhibiting 100% aqueous solubility.

6. A composition of neem leaf water-soluble bitters, the composition comprising:

a) a neem leaf hydroalcoholic extract at a concentration of 70%;
b) a polyvinyl pyrrolidone K-30 (PVP K-30) at a concentration of 27.5%; and
c) a polyoxyethylene sorbitan monooleate at a concentration of 2.5%.

7. The composition as claimed in claim 5, wherein said neem leaf water-soluble bitters comprises flavonoids at a concentration range of 8%-15%.

8. The composition as claimed in claim 5, wherein said composition inhibits the generation of reactive oxygen species with the percentage inhibition of 75%.

9. The composition as claimed in claim 5, wherein said composition inhibits the inflammatory mediator cyclooxygenase-2 (COX-2) with the percentage inhibition of 47%.

10. The composition as claimed in claim 5, wherein said composition is safe and non-toxic up to a concentration of 7.2 mg/ml.