DESC:FIELD OF THE INVENTION
[001] The invention relates to a Phyto-actives (Plant sourced API’s) composition, phytochemical and plant medicine composition and a method of manufacturing the same.

BACKGROUND OF THE INVENTION
[002] Chronic Obstructive Pulmonary Disease (COPD) is a progressive disease of the airways and pulmonary that is characterized by a gradual loss of lung function leading to death because of 60% - 100% lung dysfunction. This dysfunction often referred to as “lung fibrosis” has many causes such as, air pollution, cigarette smoke inhalation, viral/bacterial infection, allergy causing irritants (such as pollen, occupational hazardous fumes, dust, chemicals etc.) and genetic causes.

[003] The term COPD is often widely used to describe many different pulmonary conditions including Asthma, chronic bronchitis, chronic obstructive bronchitis, and emphysema. Most of which are not reversable/treatable in a chronic form. The chronic obstruction/block of lung airflow are the major symptoms in the COPD can be described as lung damage caused majorly by fibrosis leading to over 4 million / year deaths, COPD is the second largest death causing disease accounting for 5.36% of total death worldwide just behind heart disease.

[004] COPD develops due to irritants such as smoke, dust etc. which are inhaled. Chronic bacterial infections also cause inflammation. The damage of the connective tissue of the lungs leads to emphysema, which in turn causes poor airflow, poor absorption, release of respiratory gases and narrowing of the airways owing to the inflammation, fibrosis within connective lung tissues leading to the COPD.

[005] The major symptoms of COPD are chronic cough, high level of sputum production and severe shortness of breath. COPD is usually diagnosed by spirometry which is a test for the presence of airway obstruction. As per the national health service (NHS-UK), there's no cure for chronic obstructive pulmonary disease (COPD), but management medication available can help slow down the progression of the condition.

[006] Accordingly, the present treatment options for patients with COPD are generally aimed at slowing the progression of the disease. Medical treatment includes bronchodilators, such beta-2-agonists which relax the smooth muscle thereby decreasing obstruction. Anti-inflammatory agents, such as corticosteroids are often used to treat the inflamed airways of COPD sufferers. Other medical treatment options include mucolytics which act to break up the mucus that blocks the airways of patients with COPD. The major goals of management are to reduce risk factors, prevent and treat acute exacerbations, and manage associated illnesses.

[007] Another approach to manage COPD involves the use of monoclonal antibodies (MCA) therapies to manage COPD conditions. The existing two MCA’s patented products are in the market are Mepolizumab and Benralizumab.

[008] However, there are various shortcomings in the available treatments with many side effects. For instance, the main side effects of beta -2 agonists like salbutamol include trembling, particularly in the hands, nervous tension, headaches, suddenly noticeable heartbeats (palpitations), muscle cramps etc. More serious side effects are rare but can include sudden tightening of the airways (paradoxical bronchospasm) with some inhalers. Excessive doses can occasionally cause heart attacks and a severely low level of potassium in the blood (hypokalemia). For certain medications 30% of patients may not respond.

[009] Further, the main side effects of anticholinergics like ipratropium involve dry mouth, heart burn, cataract, constipation, dysphagia (Swallowing difficulty), glaucoma, cough, palpitation, tachycardia, headaches, throat Irritation, arrhythmia, nausea, difficulty in urinating, insomnia etc.

[010] Therefore, there is a need for a composition which covers maximum pathophysiology and mechanism for the management of COPD. This is achieved in the present invention by direct lung delivery formulations comprising herbs and phytopharmaceuticals for all age groups such as elderly, adult as well as children and infants.

[011] Accordingly, the present invention discloses a composition that is useful at delivering the respirable plant-based formulation directly to the lungs for its repair, regeneration, rejuvenation, re-modelling and is useful for the treatment of COPD and other lung related disorders such as asthma, bronchitis, pneumonia, pulmonary hypertension, etc.

[012] According to the present disclosure, there are four major mechanisms, which are to be managed individually and together to achieve the desired efficacy, in the treatment of COPD such as lung repair and regeneration, bronchodilation, prevention form infections and anti-microbial treatment, immunomodulation and decreased inflammation.

SUMMARY OF THE INVENTION
[013] In one aspect, the present invention provides a composition comprising composition comprising: at least one phytopharmaceutical and at least one nutraceutically or pharmaceutically acceptable excipient.

[014] In another aspect, the present invention provides a method for preparing the composition.

[015] In still another aspect, the present invention provides a method of treatment of lung diseases.

[016] In yet another aspect, the present invention relates to use of the composition in treatment of lung diseases.

BRIEF DESCRIPTION OF THE DRAWINGS
[017] Reference will be made to embodiments of the invention, examples of which may be illustrated in accompanying figures. These figures are intended to be illustrative, not limiting. Although the invention is generally described in the context of these embodiments, it should be understood that it is not intended to limit the scope of the invention to these particular embodiments.

Figure 1 shows the percent change in f (Breaths per minute) in different treatment groups.
Figure 2 shows the percent change in TVb (volume inhaled in ml) in different treatment groups.
Figure 3 shows the percent change in MVb (the rate of ventilation in mL/min) in different treatment groups.
Figure 4 shows the level of TNF-a (ng/mg protein) in the lung tissues of different treatment groups.
Figure 5 shows the level of IL-6 (pg/mg protein) in the lung tissues of different treatment groups.
Figure 6 shows the histopathological images of lung tissues of different treatment groups (Animal Study-I)
Figure 7 shows the histopathological images of lung tissues of different treatment groups (Animal Study-II)

DETAILED DESCRIPTION OF THE INVENTION
[018] The present invention discloses composition of plant actives (Plant sourced API’s), phytochemical and plant medicine composition and a method of manufacturing the same. In an aspect of the present invention disclosed is the composition exhibit efficacy for the COPD and other lung related disorders. The composition comprises herbs and phytopharmaceuticals and at least one nutraceutically or pharmaceutically acceptable excipient.

[019] An aspect of the present invention discloses a composition comprising:
a. at least one phytopharmaceutical; and
b. at least one nutraceutically or pharmaceutically acceptable excipient.

[020] In an embodiment of the present invention the plant-derived herbs and phytopharmaceuticals are selected from the group consisting of, but not limited to, scutellarin, baicalin, quercetin, eucalyptus oil, camphor oil, curcumin, dronabinol from cannabis, atropine and cordyceps or mixture thereof. Typically, the phytochemicals can be extracted from the plants belonging to the genus Erigeron, Scutellaria, Benincasa, Carissa, Vitis, Eucalyptus, Cinnamomum, Cannabis, Cordyceps, Atropa etc. respectively.

[021] In an embodiment of the present invention the plant-derived herbs and phytopharmaceuticals are selected from the extract(s), fraction(s), active compound(s) and phytochemical(s) or mixtures thereof from the group consisting of, but not limited to, scutellaria, cannabis, cordyceps, fenugreek, eucalyptus, clove, mint, CBD, licorice, camphor, godmania, tridex, andrographolide, turmeric, atropa etc. Erigeron breviscapus, Scutellaria baicalensis, Benincasa hispida, Carissa Congesta, Vitis vinifera, Eucalyptus globulus, Cinnamomum camphora, Cannabis sativa, Cannabis indica, Cordyceps militaris, Cordyceps sinensis, Atropa belladonna, Trigonella foenum, Andrographis paniculata etc. These ingredients are derived from conventional extractions procedures. These ingredients can be the extracts or enriched fractions or pure compounds or the mixtures thereof.

[022] In an embodiment of the present invention the plant-derived herbs and phytopharmaceuticals are selected from the extract(s), fraction(s), active compound(s) and phytochemical(s) or mixtures thereof from the group consisting of, but not limited to, Scutellaria baicalensis, Godmania aesculifolia, Tridex procumben, Atropa belladonna (dhatura), Erigeron breviscapus, Benincasa hispida, Carissa Congesta, Vitis vinifera, Eucalyptus globulus, Cinnamomum camphora, Cannabis sativa, Cannabis indica, Cordyceps militaris, Cordyceps sinensis, Trigonella foenum, Andrographis paniculata etc.

[023] Typically, the plant extracts are derived from plant material selected from various parts of the plant such as, but not limited to, roots, rhizomes, stems, seeds, barks, flowers, leaves and fruits and is extracted using conventional extraction techniques using conventional solvents. Accordingly, the extracts or the raw materials can be sourced from any of these different natural sources.

[024] The conventional solvents are selected from the group of, but not limited to, water, alcohol, organic solvent and a combination thereof or methods such as cryogenic extraction, maceration, infusion, decoction, percolation, hot continuous extraction (Soxhlet), aqueous alcoholic extraction, by fermentation, counter-current extraction, ultrasound extraction (sonication), cold pressed extraction and supercritical fluid extraction whichever is suitable to obtain complete extract. The extracts may be in solid form or semi-solid form or liquid form or nano-emulsion form or suspension form.

[025] The scope of the present invention is not only limited to Scutellaria, cannabis, cordyceps, fenugreek, eucalyptus, clove, mint, CBD, licorice, camphor, godmania, tridex, Andrographis , turmeric, Atropa Erigeron, , Benincasa, Carissa, Vitis, Cinnamomum, Cannabis, ,, Trigonella, etc. plants and products derived therefrom but also extends to botanically closely related plants specially belonging to same family, preferably belonging to same genus, still preferably belonging to same species having substantially similar phenotypic and genotypic characteristics.
[026] Typically, in the composition of the present invention, the amount of Scutellarin can be in the range of 1 to 75 wt %, the amount of Baicalin can be in the range of 1 to 80 wt %, the amount of Quercetin can be in the range of 1 to 60 wt %, the amount of Eucalyptus oil can be in the range of 0.1 to 10 wt %, the amount of Camphor oil can be in the range of 0.1 to 10 wt %, the amount of Curcumin can be in the range of 0.1 to 20 wt %, the amount of Dronabinol from Cannabis can be in the range of 0.1 to 25 wt %, the amount of Atropine can be in the range of 0.1 to 50 wt %, the amount of Cordyceps can be in the range of 0.1 to 40 wt %, and the amount of at least one nutraceutically or pharmaceutically acceptable excipient can be in the range of 0.1 to 30 wt %.

[027] In a preferred embodiment, the composition comprises 0 to 97 wt% of Scutellarin, Baicalin, and Quercetin, and 0 to 3 wt% of Camphor oil.

[028] In another preferred embodiment, the composition comprises 0 to 90 wt% of Scutellarin, Baicalin, and Quercetin, and 0 to 10 wt% of Camphor Oil, Mentha Oil and Carom Seed Oil.

[029] In one of the embodiments, the active ingredients having micro and nano sizes. Typically, up to 400 mm particle size or nano molecules are preferably used for the composition.

[030] In one of the embodiments, without wishing to be bound by any theory, the active ingredients of the composition, but not limited to, are as follows:
Plant Active ingredient
Scutellaria baicalensis
Scutellarin
Baicalin
Quercetin Vegetable sources like onion
Cannabis Dronabinol
Cordyceps Cordycepin
Eucalyptus/Clove/Mint/CBD/Licorice/oil/Camphor etc. _
Fenugreek Trigonelline, 4-Hydrox-isolecucine
Godmania aesculifolia, Tridex Procumben
7,8 Dihydroxy Flavones
Andrographis Andrographolide
Curcuma longa (Turmeric) Curcumin
Curcuminoids
Atropa belladonna (Dhatura) Atropine

[031] In an embodiment of the present invention, at least one nutraceutically or pharmaceutically acceptable excipient is selected from the group consisting of, but not limited to, at least one diluent, at least one super disintegrant, at least one binder, at least one lubricant, at least one glidant and combinations thereof. Typically, the at least one nutraceutically or pharmaceutically acceptable excipient is selected from the group consisting of, but not limited to, Lactose monohydrate, Mannitol, Trehalose, L-leucine, HPMC, Cellulose, Chlorofluorocarbons, Fluorocarbons (trichloromonofluoromethane, dichlorodifluoromethane), Saline, Ethanol, HFC-134a (Hydrofluorocarbon), HFC-227 hydrocarbons (propane, butane, isobutane), Tetrafluoroethane, Ethanol, (CF3CH2F) Hydrochlorofluorocarbons, inert gases (nitrogen, NO2, CO2), Compressed gases, Sorbitol, Mannitol, Polyethylene glycol, Acetic acid, Sodium bicarbonate, Glycerine etc.

[032] The composition of the present invention may be in the form of, but not limited to, a semi-solid mass, powder, an oil and water-soluble dispersion, nano emulsion, a capsule, tablet, syrup, a blend, a suspension, nasal drop or spray, dry powder, granules, micronized dry powder, respirable fluid/aerosols and the like. In an exemplary embodiment of the present invention the composition of the present invention can be encapsulated and in a dosage form of a capsule. Depending upon the dosage the composition may comprise further excipients necessary for the manufacture of the preferred dosage form and its breakdown following ingestion. In another embodiment of the present invention, the composition may further comprise Lactose monohydrate, Mannitol, Trehalose, L-leucine, HPMC, Cellulose, Chlorofluorocarbons, Fluorocarbons (trichloromonofluoromethane, dichlorodifluoromethane), Saline, Ethanol, HFC-134a (Hydrofluorocarbon), HFC-227 hydrocarbons, Propane, Butane, Isobutane), Tetrafluoroethene, Ethanol, (CF3CH2F)Hydrochlorofluorocarbons, inert gases (nitrogen, NO2, CO2), Compressed gases, Sorbitol, Mannitol, Polyethylene glycol, Acetic acid, Sodium bicarbonate, Glycerin etc.

[033] Advantageously, the composition of the present invention exhibits the below mentioned properties:
- Repairs and regenerates lung to normalize the lung functions such as bronchodilation, immunomodulation and proper breathing pattern by modulation of PI3K/AKT/NF-kB pathway, reversing pulmonary fibrosis through inhibiting NF-?B/NLRP3-mediated epithelial–mesenchymal transition and inflammation, Secretoglobin family 1a member 1 (SCGB1A1) and club cells-based lung injuries repairing pathways, using SIRT1 gene downregulation, epigenetically regulating the expression of important genes by reversing DNA methylation and altering histone modifications and by targeting several miRNA that play a key role in diseases like COPD, ALI (Acute Lung Injuries), ARDS (Acute Respiratory Distress Syndrome), IPF (Idiopathic Pulmonary Fibrosis).
- Decrease pulmonary obstruction by increasing FEV1 (Forced Expiratory Volume in 1 sec). Accordingly, to improve the FEV1 value minimum of 10% & more. The standard drug i.e., Salbutamol improves FEV1 value in the range of 4%-6%.
- Acts as a the potent antibacterial in order to prevent the inflammation/immune hyperreaction in COPD subjects.
- Achieves a minimum of 20% improvement in the immunomodulatory response of animal model & human subjects.
- Decreases inflammation which is observed from inflammation markers. It is observed that inflammation markers will be reduced and minimize up to 20% & more.
- Achieves minimum of 10% improvement in the breath pattern of COPD subjects and walk test.

[034] In another aspect the present invention discloses method for manufacturing the Phyto-actives (Plant sourced API’s) composition, phytochemical, and plant medicine composition of the present invention. Typically, the method involves steps such as 1) in process quality control checks of the individual ingredients 2) conversion of macro molecules to the nano/micro sizing by different methodology such as solvent/antisolvent methods, sonication, high speed centrifugation, ultrafiltration, sieving with 200 to 800 numbers sieves etc. 3) Mixing of individual ingredients in the optimized ratio. 4) Addition of excipients in the optimized ratio 5) In-vitro quality checks for the final formulation such as Anderson cascade impactor analysis (ACIA) 6) Conversion of the final combination in the Dry Powder Inhalable form/Metered Dose Inhaler/Respiratory fluids/Nasal Spray/Oral solid dosage/Syrup/Dermal etc. 7) Efficacy study of final formulation in COPD/lung disorders induced animal models.

[035] In a preferred embodiment, the method of preparing the composition comprising: at least one phytopharmaceutical; and at least one nutraceutically or pharmaceutically acceptable excipient, comprising the following steps:
1) extracting phytopharmaceutical from plants using solvents,
2) conversion of individual ingredient macro molecules obtained from plant extract to the nano/micro size,
3) mixing of nano/micro size individual ingredients in the optimized ratio,
4) addition of nutraceutically or pharmaceutically acceptable excipients in the optimized ratio to achieve a final formulation.

[036] In a preferred embodiment, the method preferably uses solvents that are selected from Milli Q water, alcohol, organic solvent and a combination thereof.

[037] In a preferred embodiment, the method preferably uses extraction involving cryogenic extraction, maceration, infusion, decoction, percolation, hot continuous extraction (Soxhlet), aqueous alcoholic extraction, by fermentation, counter-current extraction, ultrasound extraction (sonication), cold pressed extraction and supercritical fluid extraction.

[038] In a preferred embodiment, the method preferably uses plant extracts that are in solid form, semi-solid form, liquid form, nano-emulsion form, suspension form, preferably powdered form.

[039] In another aspect the present invention discloses a method of treatment of COPD (chronic obstructive pulmonary disease) and other lung disorders (such as asthma, emphysema, bronchitis) in humans by administering doses of the composition of Phyto-actives of the present disclosure.

[040] In an aspect of the present invention the doses of the composition in humans are administered as follows once in a day, twice in a day and thrice in a day or as per prescription by physician based on the severity of diseases.

[041] In an aspect of the present invention, the method of treatment, of COPD (chronic obstructive pulmonary disease) and other lung disorders (such as asthma, emphysema, bronchitis) in humans, repairs and regenerates lung to normalize the lung functions, decreases pulmonary obstruction by increasing FEV1, prevents the inflammation/immune hyperreaction, achieves a minimum of 20% improvement in the immunomodulatory response, decreases inflammation and achieves a minimum of 10% improvement in the breath pattern.

[042] In another aspect the present invention discloses the use of the composition of Phyto-actives preferably for the treatment of COPD (chronic obstructive pulmonary disease) and other lung disorders (such as asthma, emphysema, bronchitis Idiopathic Pulmonary Fibrosis, Pulmonary viral infections, Pulmonary bacterial infections, Pneumonia, Lung cancer, Pulmonary Edema, Pneumothorax, or Atelectasis), preferably in humans.

EXAMPLE
[043] The following examples are illustrative of the invention but not limitative of the scope thereof:

[044] Materials and Methods

[045] Materials

[046] Scutellarin was obtained from Erigeron breviscapus. It is from family Asteraceae family and found only in China, in the provinces of Guangxi, Guizhou, Hunan, Sichuan, Tibet, and Yunnan. The material was procured from a commercial source in China.

[047] Baicalin was obtained from Scutellaria baicalensis. It is from Lamiaceae family found in China, Korea, Mongolia, and Russia in the Russian Far East and Siberia. The material was procured from a commercial source in China.

[048] Quercetin was obtained from Sophora japonica. It is from Fabaceae family, native to China. The material was procured from a commercial source in China.

[049] Bleomycin (BLM), Camphor Oil, Mentha Oil and Carom Seed Oil were procured from commercial sources in India.

[050] Method for the preparation of the formulation: The ratio of all the constituents was decided after the dynamic score calculation for therapeutic properties of individual ingredients. Primarily powders – Scutellarin, Baicalin and Quercetin were selected individually or were mixed to form powdered homogenous mixture. Respective oils were added slowly to avoid any clumping, by adding volume in microliters followed by continuous shaking in vortex shaker to form a uniform mixture. The obtained mixture was sonicated using 15ml of Milli Q water to achieve the desired formulation. The formulations, after undergoing sonication, were converted into aerosol particles through the nebulizer device of size ranging from 5uM to 200 uM, which was suitable for inhalation.

[051] Example-1 (Formulation-I or F1): To the Scutellarin powder (20mg), Camphor Oil (1mg), Mentha Oil (1mg) and Carom Seed Oil (1mg) each were added slowly followed by continuous shaking in vortex shaker to form a uniform mixture. The obtained mixture was sonicated using 15ml of Milli Q water to achieve the desired formulation. The formulations, after undergoing sonication, were converted into aerosol particles through the nebulizer device of size ranging from 5uM to 200 uM, which was suitable for inhalation.

[052] Example-2 (Formulation-II or F2): To the Baicalin powder (40mg), Camphor Oil (1mg), Mentha Oil (1mg) and Carom Seed Oil (1mg) each were added slowly followed by continuous shaking in vortex shaker to form a uniform mixture. The obtained mixture was sonicated using 15ml of Milli Q water to achieve the desired formulation. The formulations, after undergoing sonication, were converted into aerosol particles through the nebulizer device of size ranging from 5uM to 200 uM, which was suitable for inhalation.

[053] Example-3 (Formulation-III or F3): To the Quercetin powder (40 mg), Camphor Oil (1mg), Mentha Oil (1mg) and Carom Seed Oil (1mg) each were added slowly followed by continuous shaking in vortex shaker to form a uniform mixture. The obtained mixture was sonicated using 15ml of Milli Q water to achieve the desired formulation. The formulations, after undergoing sonication, were converted into aerosol particles through the nebulizer device of size ranging from 5uM to 200 uM, which was suitable for inhalation.

[054] Example-4 (Formulation-IV or F4): 20 mg of Scutellarin powder, 40 mg of Baicalin and 40 mg of Quercetin were mixed to form powdered homogenous mixture. Respective oils - Camphor Oil (1mg), Mentha Oil (1mg) and Carom Seed Oil (1mg) each were added slowly followed by continuous shaking in vortex shaker to form a uniform mixture. The obtained mixture was sonicated using 15ml of Milli Q water to achieve the desired formulation. The formulations, after undergoing sonication, were converted into aerosol particles through the nebulizer device of size ranging from 5uM to 200 uM, which was suitable for inhalation.

[055] The composition based on Example 1 to 4 were then taken for further studies.

[056] Table 1 below indicates the ingredients of the formulations I to IV according to Examples 1 to 4:
Table 1:
Examples Scutellarin(mg) Baicalin
(mg) Quercetin
(mg) Camphor Oil(mg) Mentha Oil (mg) Carom
Seed Oil (mg)
Formulation
-I 20 1mg 1mg 1mg
Formulation
-II 40 1mg 1mg 1mg
Formulation
-III 40 1mg 1mg 1mg
Formulation
-IV 20 40 40 1mg 1mg 1mg

[057] Animal Study-I:

[058] SD Rat (7-8-week-old) weighting about 200-250 gm were used in the present study. Animals were maintained with a 12-hour light-dark cycle and were allowed food and water ad libitum. The animal house maintains ambient condition like temperature (22 ± 2°C) and humidity (50 ± 5%). All procedure and experimental protocols were approved by the Institutional Animal Ethics Committee (IAEC) of National Institute of Pharmaceutical Educational and Research (NIPER), Hyderabad, India, as per the committee for control and supervision of experiments on Animals (CPCSEA) guidelines of the Government of India.

[059] Table 2: Treatment Groups

[060] Animals were randomly divided into six groups (Normal Control (NC), Disease Control (DC) and Treatment Groups-T1 to T4): control group (NC), Bleomycin(BLM)-induced pulmonary fibrosis group (DC), BLM-induced pulmonary fibrosis treated with four different herbal formulation according to the present disclosure at dose of 100 mg/kg. Pulmonary fibrosis model was induced by intratracheal instillation of bleomycin at the concentration of 5.0 mg/kg under anesthesia by isoflurane. Animals in the control group received intratracheal instillation of the same amount of saline solution. In other groups, animals established with pulmonary fibrosis model treated with herbal formulation by intragastric administration once per day for 14. All the animals of different treatment were evaluated for their lung functions in Whole Body Plethysmograph on day 0 before start of Treatment and at end of treatment duration after 14 days. On day 15 all the animals were sacrificed, lung tissues then collected for further measurements.

[061] Whole Body Plethysmograph: All the animals of different treatment groups were subjected to for the evaluation of various lung functional parameters including Respiratory Rate (f), Tidal volume (TVb) and Minute volume (MVb) on day 0 (Before starting the treatment) and on day 14 at the end of the treatment duration.

[062] Level proinflammatory markers: Various proinflammatory markers like Tumour Necrosis Factor-a (TNF-a) and Interlukin-6 (IL-6) were measured in the lung tissues of different treatment groups of animals.

[063] Histopathological Evaluation: The lung tissues from different treatments groups were evaluated for histopathological examinations.

[064] Results (Animal Study-I):

[065] Respiratory Rate (f): The respiratory rate of all the animals were measure on day 0 and at the end of the study. The percent change in f (Breaths per minute) from day 1 to day 14 was calculated and summarized below Table 3.

[066] Fast shallow breathing (tachypnea) is one of several breathing difficulties experienced by pulmonary fibrosis (PF) patients. Breathing problems are common in PF because the disease causes progressive scarring in the lungs which inhibits the normal exchange of oxygen gas needed for healthy breathing and blood flow. Low blood oxygen levels lead to shortness of breath (dyspnea) and shallow breathing.

[067] Rapid or shallow breathing in PF results from the patient’s inability to take in enough oxygen. Typically, a healthy person will take between 12 to 20 breaths per minute. Tachypnea is identified when the average number of breaths exceeds 20 breaths per minute (Figure 1).

[068] In the present study also, our data are in consistent with the earlier reports on PF where the change in the f (Breaths per minute) in Bleomycin treatment groups of animals showed remarkable changes in in the f on day 14 as compared to the baseline (Day 0).

[069] Tidal volume (TVb): The tidal volume of all the animals were measure on day 0 and at the end of the study. The percent change in TVb (volume inhaled in ml) from day 1 to day 14 was calculated and summarized below Table 4.

[070] Tidal volume is the amount of air that moves in or out of the lungs with each respiratory cycle. The hallmark of obstructive lung disease is difficulty expelling air out of the lungs due to progressive airway narrowing. Since the problem in obstructive lung disease is expiratory, breathing with higher tidal volumes helps overcome airway resistance. Therefore, patients acquire a breathing pattern of deep, slow breaths to minimize the work of breathing. Similarly in the present study the TVb in the bleomycin treated group of animals were increased significantly while with treatment groups it was lower down (Figure 2).

[071] Minute volume (MVb): Minute volume (the rate of ventilation) of all the animals were measure on day 0 and at the end of the study. The percent change in MVb (the rate of ventilation in mL/min) from day 1 to day 14 was calculated and summarized below Table 5.

[072] Minute ventilation is the tidal volume times the respiratory rate, usually, 500 mL × 12 breaths/min = 6000 mL/min in humans. Increasing respiratory rate or tidal volume will increase minute ventilation. Dead space refers to airway volumes not participating in gas exchange. Anatomic dead space includes air in the mouth, trachea, and all but the smallest bronchioles (Figure 3).

[073] Level of TNF-a: The TNF-a (ng/mg protein) were measured in the lung homogenates of all the animals from various treatment groups and summarized below Table 6 and shown in Figure 4.

[074] Level of IL-6: The IL-6 (pg/mg protein) were measured in the lung homogenates of all the animals from various treatment groups and summarized below Table 7 and shown in Figure 5.

[075] Histopathological Evaluation: The perfused lungs of various treatment groups were stained with H & E staining and evaluated for lung damage. The overall gross pathological observation of lung tissues is tabulated below Table 8 and shown in Figure 6:

[076] Based on the above findings it can concluded that the all the samples were found to be effective (up 80%) against BLM-induced pulmonary fibrosis group. All the treatment groups were able reverse the damage caused by bleomycin in terms of lungs vital capacity, inflammatory markers (TNF-a and IL-6) and lung cells. However, the sample 4 were found to most effective as compared to other samples to regain lung functional capacities as comparable to Normal control group. Histopathological analysis of lung tissue was correlated with the lung performance examination (Whole Body Plethysmograph) as well as inflammatory markers (IL-6, TNF) from lung tissues.

[077] Animal Study-II:

[078] Animals were randomly divided into eleven (11) groups having 12 animal each (Table 9) and were treated with the following herbal formulations according to the present disclosure.

[079] Formulation 1 (or F1) corresponds to Example 4 of the Animal Study-I wherein only camphor oil (1mg) is used, and Formulation 2 (or F2) corresponds to the example 4 of the Animal study-1.

Table 9: Various Treatment Groups

[080] All the animals except Group I (Normal Control) were exposed to cigarette and pollution (Burning smoke of Saw dust) to create COPD. The COPD model was induced in animals by smoking of 5 cigarettes/group per day and pollution (Burning smoke of Saw dust)/day for the period of 30 days. After the exposure period, all the animals except Normal Control groups were categorized in four individual group based on disease severity such as GOLD-1, Gold-2, Gold-3 and Gold-4 as a COPD groups. All the animals of COPD groups were treated with two different Herbal formulation according to the present disclosure at dose of 120 mg/kg via inhalation route (Table 1). Animals in the control group did not receive any treatment. In other groups, animals established with COPD (All 4 stages) model were treated with herbal formulation according to the present disclosure by inhalation once per day for additional 60 days. After 60 days, animals were sacrificed, lung tissues and blood then collected for further measurements.

[081] Various Parameters evaluated:

[082] Bodyweight: Bodyweight of each animal from all groups were monitored periodically up to endpoint of the study.

[083] Elisa: IL4, IL6, IL8 and TNF-a at the endpoint of the study (90 Day)

[084] CBC (Complete blood count includes total leucocyte, monocytes, Eosinophil count, RBC count, Hemoglobin, hematocrit%, Platelet count etc.): at 30 day, and at the endpoint of the study (90 Day)

[085] Bronchoalveolar lavage fluid (BALF) analysis: At the endpoint of the study (90 Day)

[086] BALF: Total protein content at the endpoint of the study (90 Day)

[087] Plethysmography. All the animals of different treatment groups were subjected to for the evaluation of various lung functional parameters including Respiratory Rate (f), Tidal volume (TVb) and Minute volume (MVb) on day 0 (Before starting the treatment) and on day 30, 60 and on 90 days at the end of the treatment duration.

[088] Histopathology: at the end of the study point: The lung tissues from different treatments groups were evaluated for histopathological examinations.

[089] Results (Animal Study-II)

[090] Body Weight: A total 132 animals were randomly divided in 11 groups having 12 animals in each group. After a week of acclimatization, all the animals were observed for body weight during exposure period of cigarette and saw dust smoke. The total body weight gain by each group is presented in table 2. At the end of the exposure of cigarette and saw dust smoke, the animals were assigned to different groups of Gold-1,2,3 and 4 based on the body weight, mortality rate and WBP parameters as presented in Table 10.

Table 10: Body weight gain by all the animals after exposure period of cigarette and saw dust smoke
Groups Total Weight gain (gm) Group Assigned
Group 1 151.60 NC (Normal Control)
Group 2 111.48 Gold-1 +F1
Group 3 115.94 Gold-1 + F2
Group 4 133.42 Gold-3 + F1
Group 5 129.39 Gold-3 + F2
Group 6 114.78 Gold-2 + F1
Group 7 120.15 Gold-2 + F2
Group 8 93.44 DC-1 (Gold-4)
Group 9 95.90 DC-1 (Gold-4) + F1
Group 10 100.23 DC-2 (Gold-2)
Group 11 102.68 DC-2 (Gold-4) + F2

[091] Body weight of all the animals of different treatment groups were monitored for further 60 day of treatment duration. The total weight of each group after the treatment of formulation 1 and 2 is given in table 11. There was a significant death noted in both disease control (DC) groups while treatment was able to maintain the health condition with survivability. Significant weight loss was noted in the DC group due to lung damage & COPD. The treatment group seems to maintain the weight of animals post-treatment with both formulations, as mentioned in the below-mentioned Table 11:

Table 11: Total body weight gain by each treatment group after the treatment

*Total number of animals died: 2
# Total number of animals died:1

[092] Respiratory Rate (f): The respiratory rate of all the animals were measure on day 0 and at the end of the exposure period of cigarette and saw dust smoke on day 30 of the study. The percent change in f (Breaths per minute) from day 0 to day 30 was calculated and summarized below.

Table: 12: Frequency (f) of respiration in different groups of animals


Comparisons between groups were done by using one-way ANOVA followed “Bonferroni's multiple comparison post-hoc test” P-value on figure indicates significance *P<0.05, **P<0.001, ***P<0.0001 vs. NC

[093] Normal control (NC) group animals did not show any significant change in the f (frequency) throughout the study period while all other group of animals showed significant increase in the f on day 30 as compared to day 0 after the exposure period of cigarette and saw dust smoke. Animals of different treatment groups G4-F1, G4-F2, G1-F1 and G1-F2 showed significant improvement in f on day 60 and 90 as compared to day 30 (Mentioned in Fig-4). On the other hand, treatment group with formulation 2 showed significant (*P<0.05) improvement & recovery in f value from day 30 to day 90. The data obtained in f value indicates that formulation 2 recovered the lung condition to normalization up to 90% in G1, G2 and G3. While G4 with formulation 2 showed upto 80% improvement in lung post treatment.

[094] Fast shallow breathing (tachypnea) is one of several breathing difficulties experienced by pulmonary fibrosis (PF) patients. Breathing problems are common in PF because the disease causes progressive scarring in the lungs which inhibits the normal exchange of oxygen gas needed for healthy breathing and blood flow. Low blood oxygen levels lead to shortness of breath (dyspnea) and shallow breathing.

[095] Rapid or shallow breathing in PF results from the patient’s inability to take in enough oxygen. Typically, a healthy person will take between 12 to 20 breaths per minute. Tachypnea is identified when the average number of breaths exceeds 20 breaths per minute.

[096] Tidal volume (TVb): The tidal volume of all the animals were measure on day 0 (Baseline), day 30 (exposure period of cigarette and saw dust smoke), day 60 and day 90 (Treatment duration) of the study.

Table 13: Tidal volume (TVb) of in different groups of animals

Comparisons between groups were done by using one-way ANOVA followed “Tukey's multiple comparisons test post-hoc test” P-value on figure indicates significance *P<0.05, **P<0.001, ***P<0.0001 vs. NC

[097] Normal control (NC) group of animals did not show any significant change in the TVb (Tidal volume) throughout the study period while all other group of animals such as DC1 & DC2 showed significant (P<0.0001) increase in the TVb on day 30 as compared to day 0 after the exposure period of cigarette and saw dust smoke. Animals of different treatment groups G1-F2, G2-F2, G4-F1 did not showed any significant changes in TVb on throughout the treatment duration. G4-F2 and G3-F1 group of animals showed significant (P<0.05) improvement in TVb on day 90 as compared to day 30 and shown significant improvement on day 60.

[098] Tidal volume is the amount of air that moves in or out of the lungs with each respiratory cycle. The hallmark of obstructive lung disease is difficulty expelling air out of the lungs due to progressive airway narrowing. Since the problem in obstructive lung disease is expiratory, breathing with higher tidal volumes helps overcome airway resistance. Therefore, patients acquire a breathing pattern of deep, slow breaths to minimize the work of breathing. Similarly in the present study the TVb in the cigarette and saw dust smoke treated group of animals were increased significantly while with treatment groups it was lower down. The TVb values seems to be recovered well post the treatment in all group specifically with the group belongs to formulations 2. However, formulation 1 groups also shown significant recovery to the lung. That concluding the effectiveness of treatment drug for the recovery and maintained life as mentioned in table no. 13.

[099] Minute volume (MVb): Minute volume (the rate of ventilation) of all the animals were measure on day 0 (Baseline), day 30 (exposure period of cigarette and saw dust smoke), day 60 and day 90 (Treatment duration) of the study.

Table 14: MVb (the rate of ventilation in mL/min) in different treatment groups


Comparisons between groups were done by using one-way ANOVA followed “Tukey's multiple comparisons test post-hoc test” P-value on figure indicates significance *P<0.05, **P<0.001, ***P<0.0001 vs. NC

[0100] Normal control (NC) group of animals did not show any significant change in the MVb (Tidal volume) throughout the study period while all other groups animals showed significant (P<0.0001) increase in the MVb on day 30 as compared to day 0 after the exposure period of cigarette and saw dust smoke. Animals of both DC groups DC1, DC-2, showed any significant improvement in MVb throughout the study including initial 30 day and rest 60 days. Treatment group specifically with formulation 2 of animals showed significant (P<0.05) improvement in MVb throughout the 90 as compared to day 30. On the other hand, treatment group G2-F2 (P<0.001), G3-F2 (P<0.0001), G4-F1 (*P<0.001) and G4-F2 (*P<0.001) showed significant improvement in MVb on day 60 and G2-F2, G3-F2, G4-F1 and G4-F2 show significant (P<0.0001) improvement in MVb on day 90 of the treatment. Mentioned in table no. 14. The results for MVb concluding the reverse of damage in the treatment group.

[0101] Minute ventilation is the tidal volume times the respiratory rate, usually, 500 mL × 12 breaths/min = 6000 mL/min in humans. Increasing respiratory rate or tidal volume will increase minute ventilation. Dead space refers to airway volumes not participating in gas exchange. Anatomic dead space includes air in the mouth, trachea, and all but the smallest bronchioles.

[0102] Level of TNF-a: The TNF-a (pg/mg protein) were measured in the lung homogenates of all the animals from various treatment groups and summarized below.
Table 15: TNF-a level in various treatment groups at the end of the study

[0103] TNF-a level were significantly increased in both DC-1 and DC-2 while DC-2 groups of animals have shown more significant (P<0.0001) as compared to NC group of animals. G4-F1 & G4-F2 showed significant decreased in TNF-a level as compared to DC-2 groups, while other treatment group were also able to reduce the TNF-a as compared to DC-2 groups. The tumor necrosis factor is the important marker to detect the degenerating condition & inflammation inside the lung. The data obtained from table 6 supporting the data from plethysmography that treatment is effectively working on the regenerating the lung disorders induced by saw dust smoke and cigarettes smoke.

[0104] Level of IL-6: The IL-6 (pg/mg protein) were measured in the lung homogenates of all the animals from various treatment groups and summarized below.

Table 16: IL-6 level in various treatment groups at the end of the study

[0105] IL-6 level were significantly increased in both DC-1 (P<0.001) and DC-2 (P<0.0001) groups of animals as compared to NC group of animals. Th treatment groups showed significant reduction in IL-6 such as G4-F1 (P<0.05), G4-F2, G3-F1 (P<0.001), G3-F2 (P<0.0001), G2-F1, G2-F2 (P<0.001) and G1-F1 (P<0.05) as compared to DC-2 groups.

[0106] Level of IL-4: The IL-4 (pg/mg protein) were measured in the lung homogenates of all the animals from various treatment groups and summarized below.

Table 17: IL-4 level in various treatment groups at the end of the study

[0107] IL-4 level were significantly increased in both DC-1 (P<0.0001) and DC-2 (P<0.05) while as compared to NC group of animals. There were significant improvement in IL-4 in treatment groups noted such G4-F2 (P<0.001), G3-F2 (P<0.0001) and G2-F2 (P<0.005) as compared to DC-2 groups of animals. The indicated that enhanced IL-4 biomarker get reversed to the normal control group mentioned in table no. 17.

[0108] CBC: Detailed haematological parameters in various treatment groups are provided below:

Table 18: WBC count

[0109] WBC level were found to be in normal range on 0 day while it significantly (P<0.0001) increased on 30 and 90 days in both DC-1 and DC-2 as compared to 0 day. All the treatment groups had shown significant (P<0.0001) increased in WBC levels on 30 days except treatment groups G4-F2, G3-F2 (P<0.001), G3-F1 (P<0.01), G1-F1 and G1-F2 (P<0.05) as compared to 0 Day. On day 90 treatment groups G4-F1, G4-F2 (P<0.001), G1-F1 (P<0.05) and G1-F2 (P<0.01) able normalize the raised WBC level significantly as compared to 30 Day.

Table 19: RBC count

[0110] RBC level were found to be in normal range on 30 and 90 days in normal control group as compared to 0 day. All the other treatment group of animals showed various significant level (P<0.0001), (P<0.001), (P<0.05) and (P<0.01) on day 30 as compared to 0 day. However, all the treatments didn’t show any change in the WBC level on 90 day as compared to 30 Day.

Table 20: HBG Level

[0111] The HBG level of all the animals from different groups was found to be in normal range on 0 day. While on 30 day, all the animals of various treatment groups had shown significant improvement except DC-1, G4-F2 and G1-F1 group of animals as compared to 0 day. However, all the animals of different treatment groups didn’t show any significant changes in HBG level on 90 day as compared to 30 day of treatment.

Table 21: HCT level

[0112] The HCT level in all the treatment groups were found to be in normal range on 0 day. Additionally, none of the treatment group had shown any significant changes in HCT level on 30 day as compared to 0 day. While on 90 day only G3-F2 treatment group had shown significant (P<0.01) increased in HCT as compared to 30 day.

Table 22: MCH level

[0113] The MCH level in all the treatment groups were found to be in normal range on 0 day. Additionally, none of the treatment group had shown any significant changes in MCH level on 30 day as compared to 0 day. While on 90 day only DC-I treatment group had shown significant (P<0.01) decrease in MCH changes as compared to 30 day.

Table 23: PLT level

[0114] The PLT level in all the treatment groups were found to be in normal range on 0 day. Additionally, none of the treatment group had shown any significant changes in PLT level on 30 day as compared to 0 day except NC (P<0.01) group of animals. While on 90 day only NC and DC-I treatment group had shown significant (P<0.01) decrease in PLT changes as compared to 30 day.

Table 24: LYM level

[0115] The % LYM level in all the treatment groups were found to be in normal range on 0 day. While all the treatment group had shown significant changes in % LYM level on 30 day as compared to 0 day except G1-F2 (P<0.01) group of animals. While on 90-day DC-I and DC-2 treatment group had shown significant (P<0.001) increase in % LYM as compared to 30 day. On the other hand, all the treatment groups were able to normalize the % LYM level on 90 day as compared to 30 day except G3-F2 and G1-F1 treatment group.

Table 25: MONO level

[0116] The % MONO level in all the treatment groups were found to be in normal range on 0 day. On 30 day all the treatment group had shown significant increase in the % MONO as compared to 0 day. While on 90-day G4-F2, G3-F2, G2-F1, G2-F2 and G1-F2 treatment group had shown significant reduction in the % MONO as compared to 30 day.

Table 26: GRAN level

[0117] The % GRAN level in all the treatment groups were found to be in normal range on 0 day. On 30 all the treatment group had shown significant changes in % GRAN as compared to 0 day except NC, DC-I, G4-F2, G1-F1 and G2-F2. While on 90-day DC-I and G4-F2 treatment group had shown significant increase in % MONO as compared to 30 day.

Table 27: PCT level

[0118] The PCT level in all the treatment groups were found to be in normal range on 0 day. While all the treatment group had shown significant changes in PCT level on 30 day as compared to 0 day except NC, DC-II and G1-F2 group of animals. While on 90-day none of the treatment group had shown any significant change in PCT as compared to 30 day.

Table 28: MPV level

[0119] The MPV level in all the treatment groups were found to be in normal range on 0 day. While all the treatment group had shown significant changes in MPV level on 30 day as compared to 0 day except NC and DC-I group of animals. While on 90-day none of the treatment group had shown any significant change in MPV as compared to 30 day except DC-I treatment group.

Table 29: PDW level

[0120] The PDW level in all the treatment groups were found to be in normal range on 0 day. While all the treatment group had shown significant increase in PDW level on 30 day as compared to 0 day. While on 90-day none of the treatment group had shown significant decrease in PDW level as compared to 30 day.

[0121] BALF: The BALF content of representative animals from each group were assessed for total leucocytes, differential count of neutrophils and lymphocytes at the end of the study. The values are as shown in table 30 below.
Table 30:

[0122] Animals of DC-I and II showed significant increase in total Leucocytes count, Neutrophils and Lymphocytes as compared to NC group of animals. All the treatment groups have shown to normalize the Leucocytes count, Neutrophils and Lymphocytes values significantly as compared to DC-I group of animals except G4-F1, G4-F2, G1-F1 and G1-F2 treatment group of animals.

[0123] Total Protein: The total protein content of the BALF fluid from representative animal of each group was analysed and presented in table below:

Table 31

Note: n=2

[0124] Histopathological Evaluation: The perfused lungs of various treatment groups were stained with H & E staining and evaluated for lung damage. The overall gross pathological observation of lung tissues is tabulated below and shown in Figure 7:

Table 32

Overall Grade score as- NAD =No Abnormality Detected, Minimal changes (+1), Mild changes (+2), Mild-Moderate changes (+3), Moderate changes (+4). Severe changes (+5).

[0125] Based on the result obtained in the present study where COPD was induced in all the animals except normal control by smoking of 5 cigarettes/group per day and pollution (Burning smoke of Saw dust)/day for the period of 30 days confirm the induction of damaged to the lungs in animals. After the exposure period, all the animals except Normal Control groups were categorized in four individual group based on disease severity such as GOLD-1, Gold-2, Gold-3 and Gold-4 as a COPD groups. All the animals of COPD groups were treated with two different formulations at dose of 120 mg/kg via inhalation route (Table 1).

[0126] Animals in the control group did not receive any treatment. In other groups, animals established with COPD (All 4 stages) model were treated with herbal formulation by inhalation once per day for additional 60 days. After 60 days, animals were sacrificed, lung tissues and blood then collected for further measurements.

[0127] Based on all the observed parameters like Bodyweight, Elisa: (IL4, IL6, IL8 and TNF-a), CBC, BALF (total leucocytes, differential count of neutrophils and lymphocytes), BALF (Total protein content). Whole Body Plethysmograph (Respiratory Rate (f), Tidal volume (TVb) and Minute volume (MVb)) and Histopathology it can be concluded that both the formulations (F1 and F2) is effective in reversing the damaged induced by cigarettes and pollution (Burning smoke of Saw dust) in all the GOLD level. The G4 and G3 and G1 level of animals treated with F1 and F2 were found to be less effective in treating COPD induced by cigarettes and pollution (Burning smoke of Saw dust) and the most effective combination was found to be G2-F2.

[0128] The foregoing description of the invention has been set merely to illustrate the invention and is not intended to be limiting. Since the modifications of the disclosed embodiments incorporating the spirit and substance of the invention may occur to the person skilled in the art, the invention should be construed to include everything within the scope of the disclosure.
,CLAIMS:We Claim:
1. A composition comprising:
a. at least one phytopharmaceutical, and
b. at least one nutraceutically or pharmaceutically acceptable excipient,
wherein the phytopharmaceuticals are selected from the group consisting of scutellarin, baicalin, quercetin, eucalyptus oil, camphor oil, curcumin, dronabinol from cannabis, atropine and cordyceps or mixture thereof.
2. The composition as claimed in claim 1, wherein the phytopharmaceuticals are extracted from the plants belonging to the genus Erigeron, Scutellaria, Benincasa, Carissa, Vitis, Eucalyptus, Cinnamomum, Cannabis, Cordyceps, Atropa.
3. The composition as claimed in claim 1, wherein the amount of at least one phytopharmaceutical is selected from
- 1 to 75 wt % of Scutellarin,
- 1 to 80 wt % Baicalin,
- 1 to 60 wt % Quercetin,
- 0.1 to 10 wt % of Eucalyptus oil,
- 0.1 to 2 wt % of Camphor oil,
- 0.1 to 20 wt % of Curcumin,
- 0.1 to 25 wt % of Dronabinol from Cannabis,
- 0.1 to 50 wt % of Atropine,
- 0.1 to 40 wt % of Cordyceps, and
- 0.1 to 30 wt % at least one nutraceutically or pharmaceutically acceptable excipient.
4. The composition as claimed in claim 1, wherein the composition comprises
- 0 to 97 wt% of Scutellarin, Bacailin, and Quercetine, and
- 0 to 3 wt% of Camphor oil.
5. The composition as claimed in claim 1, wherein the composition comprises
- 0 to 90 wt% of Scutellarin, Bacailin, and Quercetine, and
- 0 to 10 wt% of Camphor Oil, Mentha Oil and Carom Seed Oil.
6. The composition as claimed in claim 1 to 5, wherein at least one nutraceutically or pharmaceutically acceptable excipient is selected from a diluent, a super disintegrant, a binder, a lubricant, a glidant and combinations thereof.
7. The composition as claimed in claim 1 to 6, wherein at least one nutraceutically or pharmaceutically acceptable excipient is selected from Lactose monohydrate, Mannitol, Trehalose, L-leucine, HPMC, Cellulose, Chlorofluorocarbons, Fluorocarbons (trichloromonofluoromethane, dichlorodifluoromethane), Saline, Ethanol, HFC-5 134a (Hydrofluorocarbon), HFC-227 eahydrocarbons (propane, butane, isobutane), Tetrafluoroethane, Ethanol, (CF3CH2F) Hydrochlorofluorocarbons, inert gases (nitrogen, NO2, CO2), Compressed gases, Sorbitol, Mannitol, Polyethylene glycol, Acetic acid, Sodium bicarbonate and Glycerine.
8. The composition as claimed in claim 1 to 7, wherein the composition is in the form of a semi-solid mass, an oil and water-soluble dispersion, nano particles, a nano emulsion, a capsule, a tablet, a syrup, a blend, a suspension, a nasal drop or spray, granules, respirable fluid/aerosols, or powders.
9. The composition as claimed in claim 1 to 8, wherein the composition is in the form of a powder preferably a dry powder or a micronized dry powder.
10. A method of preparing the composition comprising: at least one phytopharmaceutical; and at least one nutraceutically or pharmaceutically acceptable excipient, comprising the following steps:
1) extracting phytopharmaceutical from plants using solvents,
2) conversion of individual ingredient macro molecules obtained from plant extract to the nano/micro size,
3) mixing of nano/micro size individual ingredients in the optimized ratio,
4) addition of nutraceutically or pharmaceutically acceptable excipients in the optimized ratio to achieve a final formulation.
11. The method as claimed in claim 10, wherein the nano/micro sizing is done by solvent/antisolvent methods, sonication, high speed centrifugation, ultrafiltration, sieving with 200 to 800 numbers sieves.
12. The method as claimed in claim 10, wherein the dosage form is selected from Dry Powder Inhalable form, Metered Dose Inhaler, Respiratory fluids, Nasal Spray, Oral solid dosage, Syrup, Dermal.
13. The method as claimed in claim 10, wherein the solvents are selected from Milli Q water, alcohol, organic solvent and a combination thereof
14. The method as claimed in claim 10, wherein the extraction involves cryogenic extraction, maceration, infusion, decoction, percolation, hot continuous extraction (soxhlet), aqueous alcoholic extraction, by fermentation, counter-current extraction, ultrasound extraction (sonication), cold pressed extraction and supercritical fluid extraction.
15. The method as claimed in claim 10, wherein the plant extracts is in solid form, semi-solid form, liquid form, nano-emulsion form, suspension form.
16. A method of treatment of lung diseases wherein the method involves administering doses of the composition as claimed in in claim 1 to 9.
17. The method as claimed in claim 16 wherein the lung diseases are preferably selected from chronic obstructive pulmonary disease (COPD), asthma, emphysema and bronchitis.
18. The method as claimed in claim 16 to 17 wherein the doses are administered as follows once in a day, twice in a day or thrice in a day.
19. Use of the composition as claimed in claims 1 to 9.
20. Use of composition as claimed in claims 1 to 9 for the treatment of chronic obstructive pulmonary disease (COPD).
21. Use of composition as claimed in claims 1 to 9 for the treatment of asthma, emphysema, bronchitis, Idiopathic Pulmonary Fibrosis, Pulmonary viral infections, Pulmonary bacterial infections, Pneumonia, Lung cancer, Pulmonary Edema, Pneumothorax, or Atelectasis.
Dated this 20th day of January 2022.
NBI Biosciences Pvt Ltd
By their Agent & Attorney

(Adheesh Nargolkar)
of Khaitan & Co
Reg. No. IN/PA-1086