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: – Polyherbal Formulation for management
of non-alcoholic fatty liver diseases (NAFLD)
2. APPLICANT:
a) NAME : Shripad Medi-Search Pvt Ltd
b) NATIONALITY : Indian Company
c) ADDRESS : 32/2A, Erandwane, Gulwani Maharaj Road, Pune
411004, Maharashtra, India.
3. PREAMBLE TO THE DESCRIPTION
The following specification particularly describes the invention and the manner in which it
is to be performed.
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Polyherbal Formulation for management of non-alcoholic fatty liver diseases (NAFLD)
Field of The Invention
Non-alcoholic fatty liver diseases (NAFLD) is a term used for range of diseases 5 wherein fat is accumulated in the liver. NAFLD is characterized by fatty infiltration of the liver, steatosis, steatohepatitis, and cirrhosis. Currently, there is no FDA approved medicine for treating fatty liver. Present invention provides Polyherbal / Ayurvedic Formulation for treating fatty liver. The Ayurvedic (polyherbal) formulation of the present invention contains Kirat- tikta (Swertia chirata), Katuka 10 (Picrorrhiza kurroa), Lashun (Alium sativum), Jiraka (Cuminum cyminum), Shunthi (Zingiber officinalis), Pippali (Piper longum), Marich (Piper nigrum), Ramatha (Asafoetida), Jasad bhasm (Zinc oxide) and Shankh bhasm (Incinerated Conch shell) in 1-4 compositions. Single composition has all 11 ingredients. Each of the 2-4 compositions contain from 2 – 8 ayurvedic medicines in lower amounts 15 enabling them to accommodate in unit formulation such as a tablet or a capsule or a sachet.
Object of the invention
The first object of the invention is to provide an Ayurvedic formulation for treating 20 fatty liver diseases, more particularly, Non-alcoholic fatty liver diseases. Ayurvedic formulation of the present invention relieves patients from elevated levels of one or more of LDL, triglycerides, blood glucose, HbA1c, bilirubin (total and direct), alanine transaminase (ALT) and aspartate transaminase (AST), alkaline phosphatase (ALP). The Ayurvedic formulation of the present invention also helps 25 in enhancing levels of one or more anti-oxidant enzymes selected from Superoxide dismutase, catalase, malondialdehyde, plasma glutathione peroxidase (table 7). The enhancement in the levels of anti-oxidant enzymes indicate readiness for the catalytic removal of free radicals and reactive species.
The second object of the invention is to provide an Ayurvedic formulation which 30 can accommodate several Ayurvedic medicines in lower amounts so that the
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medicines can act synergistically to produce effect and can be provided in a unit formulation such as a tablet or a capsule or a sachet.
Alternatively, liquid formulations such as syrup, suspensions or any other oral delivery form can be employed.
Third object of the invention is to provide an Ayurvedic formulation for treating 5 fatty liver diseases wherein the Ayurvedic formulation contains a single composition or 2-4 separate compositions, each containing 2 – 8 Ayurvedic medicines.
Fourth object of the invention is to provide a kit containing 2 - 4 separate compositions each containing 2 – 8 Ayurvedic medicines. 10
Fifth object of the invention is to provide a process to prepare an Ayurvedic formulation for treating fatty liver diseases wherein Ayurvedic formulation can accommodate several Ayurvedic medicines in lower amounts so that the medicines can act synergistically to produce effect and can be provided in a unit formulation such as a tablet or a capsule or a sachet. Alternatively, liquid formulations such as 15 syrup, suspensions or any other oral delivery form can be employed.
Background of the Invention
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M. Karan et al evaluated the methanol extract of Swertia chirata for antihepatotoxic activity against carbon tetrachloride induced liver toxicity in experimental rats. The extract was found to be active and on fractionation into butanol soluble and chloroform soluble fractions, the activity was traced and found more profound in the chloroform soluble fraction. The butanol soluble bitter rich fraction showed 25 marginal activity.
Kaushal, K. et al demonstrated that hydroalcoholic extract of S. chirata was a potent hepatoprotective intervention which was associated with its potential to alleviate oxidative stress and improve liver functions. Moreover, it could find clinical application as a safer and alternative remedy for liver ailments. 30
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However, till date no Ayurvedic medicine is prepared from Swertia chirata for daily use to treat Non-alcoholic fatty liver diseases. Swertia chirata Mother Tincture by Dr. Willmar Schwabe India Pvt. Ltd. is available for both acute and chronic malarial fevers. Chirata extract by BIZPRESSIONS.COM is available and recommended for hair Oil, Cold Press Soap, Melt and Pour Soap, Bath Bomb, 5 Lotion, Salt & Scrubs, Lip Balms, and many more cosmetic products.
Sanaz Pourreza et al has reviewed Therapeutic Effects and Mechanisms of Action of Garlic (Allium sativum) on Nonalcoholic Fatty Liver Disease in the review article published in 2022 in the “Hindawi”. It concludes as follows,
Overall, the beneficial effects of garlic in the treatment of NAFLD make it a 10 potential therapeutic and efficient agent in managing NAFLD and its related risk factors. “There is an insufficient number of clinical trials addressing the effects of garlic in humans; therefore, conducting more human research in the future is recommended”.
This review article mentions quite high doses of garlic which can vary from 400 mg 15 to 4gm per day. If such high doses are employed, it has to be given alone and cannot be combined with other herbal medicines to administer it in unit formulation / unit dosage form.
Picrorrhiza kurroa, commonly known as Katuki or Kutki has multiple uses. It is reported for balancing Kapha and Pitta. It has digestive, anti-diabetic, anti-20 asthmatic, anti-inflamatory, immune modulatory, hypolipemic and hepatoprotective activities. Its minimum dose is 500 mg daily which can go to as high as 3 – 4 g per day for chronic fever.
Although prior arts mention hepatoprotective uses or use for hepatic disorders of several Ayurvedic medicines, their daily dosage are in grams. If these dosages are 25 reduced to not more than 500 mg daily, preferably not more than 400 mg daily or more preferably below 250 mg / composition and between 1 – 200 mg / composition, there is no evidence that an effective cure can be provided.
There is no art which can teach multiple for example, how 2 – 8 Ayurvedic medicines each of which is required in large daily dose amounts can be packed in a 30
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unit formulation / unit dose and such unique formulation / dose can be administered to treat Non-alcoholic fatty liver diseases (NAFLD ).
Summary of the Invention
Under the first aspect, the invention provides an Ayurvedic formulation for treating 5 fatty liver diseases, particularly, Non-alcoholic fatty liver diseases (NAFLD).
Ayurvedic formulation of the present invention relieves patients from elevated levels of one or more of LDL, triglycerides, blood glucose, HbA1c, bilirubin (total and / or direct), alanine transaminase (ALT) and aspartate transaminase (AST), alkaline phosphatase (ALP). The Ayurvedic formulation of the present invention 10 also helps in enhancing levels of one or more anti-oxidant enzymes selected from Superoxide dismutase, catalase, malondialdehyde, plasma glutathione peroxidase (table 7). The enhancement in the levels of anti-oxidant enzymes indicate readiness for the catalytic removal of free radicals and reactive species by the body.
While elevated levels of several parameters are controlled and levels of anti-oxidant 15 enzymes are enhanced, the ayurvedic formulation does not affect hematological parameters. Levels of hemoglobin, PCV, WBC-DLC, MCV, MCH, MCHC, MPV etc. are unaffected.
Under the second aspect, the invention provides an Ayurvedic formulation which can accommodate several Ayurvedic medicines in lower amounts so that the 20 medicines can act synergistically to produce effect and can be provided in a unit formulation such as a tablet or a capsule or a sachet. The ayurvedic medicines employed are selected from herbal medicines and mineral medicines. The herbal medicines include Kirat- tikta (Swertia chirata), Katuka (Picrorrhiza kurroa), Lashun (Alium sativum), Jiraka (Cuminum cyminum), Shunthi (Zingiber 25 officinalis), Pippali (Piper longum), Marich (Piper nigrum), Ramatha (Asafoetida). The mineral medicines include Jasad bhasm (Zinc oxide) and Shankh bhasm (Incinerated Conch shell).
All herbal and mineral medicines are employed in much lower amounts than their usually reported ranges. Preferably, content of each medicine in each composition 30
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not more than 1 g, preferably not more than 500 mg and more preferably not more than 400 mg in daily dosage regime and most preferably not more than 250 mg / composition.
Preferably, each herbal and mineral medicine is employed in amounts from 1 mg – 500 mg / composition, preferably in amounts from 1 mg – 250 mg, more preferably 5 from 5 mg – 225 mg/ composition, most preferably from 10 mg – 200 mg/ composition.
Thus, if minimum 1 mg and maximum of 500 mg of an ingredient is contained in a capsule formulation containing 1000 mg fill weight, the % of the ingredient is minimum of 0.1 % to 50 %, preferably from 0.1 % to 25 %, more preferably from 10 0.5 % to 22.5 % and most preferably, from 1 % to 20 % of the weight of formulation.
The final blend or blends containing eleven ingredients ready for encapsulation into single capsule or 2-4 capsules is considered as 1 single formulation. For convenience while calculating percentages, capsule weight is not taken into consideration but total fill weight having all eleven ingredients is considered as 15 formulation.
Under the third aspect, the invention provides an Ayurvedic formulation for treating fatty liver diseases wherein the Ayurvedic formulation contains a single composition or 2 – 4 separate compositions, each containing 3 – 8 Ayurvedic medicines. 20
Fourth aspect of the invention is to provide a kit containing 2 – 4 separate compositions, each containing 3 – 8 Ayurvedic medicines.
Fifth aspect of the invention is to provide a process to prepare an Ayurvedic formulation for treating fatty liver diseases wherein Ayurvedic formulation can accommodate several Ayurvedic medicines in lower amounts so that the medicines 25 can act synergistically to produce effect and can be provided in a unit formulation such as a tablet or a capsule or a sachet.
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Brief Description of Figures
Figure 1: Schematic showing mechanism of action of herbal medicine in treatment of NAFLD.
Figure 2 shows the study flow diagram of the double blind study.
Figure 3: Column chart showing mean MRI findings at baseline and day 90. 5
Figure 4: Column chart showing number of patients according to USG grades at day 90 in two groups.
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Detailed Description of the invention
The present invention provides an Ayurvedic / polyherbal formulation for treating Non-alcoholic fatty liver diseases (NAFLD ) which is a term used for range of diseases wherein fat is accumulated in the liver. NAFLD is characterized by fatty infiltration of the liver, steatosis, steatohepatitis, and cirrhosis. 5
The ayurvedic medicines employed in the formulation of the present invention are selected from herbal medicines and mineral medicines. The herbal medicines include Kirat- tikta (Swertia chirata), Katuka (Picrorrhiza kurroa), Lashun (Alium sativum), Jiraka (Cuminum cyminum), Shunthi (Zingiber officinalis), Pippali (Piper longum), Marich (Piper nigrum), Ramatha (Asafoetida). The mineral 10 medicines include Jasad bhasm (Zinc oxide) and Shankh bhasm (Incinerated Conch shell).
These medicines are chosen based on knowledge of their biological activities reported below:
Kirat- tikta (Swertia chirata) 15
The varied ethnobotanical uses of S. chirayita have led to the initiation of various pharmacological investigations. Previous research demonstrates that the S. chirayita extracts exhibit a wide range of biological activities, such as antibacterial, antifungal, antiviral, anticancer, anti-inflammatory, and others like antidiabetic and antioxidant activities. 20
Anti-hepatitis B virus activity of S. chirayita extracts was studied on HepG cells line. The whole plant of S. chirayita has been reported for the anti-inflammatory and hypoglycemic activity.
Katuka (Picrorrhiza kurroa)
Intervention with standardized plant extracts of P. kurroa regressed several features 25 of NAFLD like lipid content of the liver tissue, morphological regression of fatty infiltration, hypolipidemic activity, and reduction of cholestatis. The present study
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has led to drug development based on reverse pharmacology for management of NAFLD.
Jasad bhasm (oxide of zinc)
Zinc is an indispensable inorganic element universally used in medicine, biology, and industry. Also, it is an essential constituent of bones, teeth, enzymes, and many 5 functional proteins.
Zinc oxide nanoparticles and zinc nanoparticles coated with soluble polymeric material may be used for treating wounds, ulcers, and many microbial infections besides being used as drug carrier in cancer therapy.
It has great potential as a safe antibacterial drug which may replace antibiotics in 10 future.
Shankh bhasm (calx of conch shell)
Rasashastra texts have mentioned Shankha Bhasma/incinerated conch (calcite form) for internal use in several diseases.
Lashun (Alium sativum) 15
Garlic showed a satisfactory effect as a hypertensive remedy by regulating high cholesterol levels and stimulating the immune system. Garlic has shown promising lipid-lowering effects on hyperlipidemic patients through the reduction of serum cholesterol concentration.
In diabetic patients the combination of garlic with olive oil effectively regulated 20 serum cholesterol and triglycerides levels, as well as dyslipidemia. Garlic in non-powder and powder forms undoubtedly reduced serum lipids levels over a one to three-month period. After four months, the consumption of GE raised HDL and lowered LDL and cholesterol levels in 23 hyperlipidemic patients.
Several studies showed that the administration of aged black garlic or garlic tablet 25 at a dose 300 mg or 6 g two times per day for 4 or 12 weeks reduced the levels of total cholesterol (TC), triglycerides, and LDL while it elevated that of HDL in patients with mild hypercholesterolemia and dyslipidemia or type 2 diabetics.
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Moreover, aged garlic reduced TC and TG at a dose of 2.4 g per day for two weeks in patients with coronary artery disease.
Raw crushed garlic at 100 mg/kg two times per day, that reduced SBP and DBP via downregulation of TG level and upregulation of serum HDL cholesterol after four weeks’ consumption in patients with metabolic syndrome. 5
Jiraka (Cuminum cyminum)
Cumin and caraway products (oils as well as their aqueous and solvent derived extracts) have shown significant antioxidant activity in several test methods which has been attributed largely to the presence of monoterpene alcohols, linalool, carvacrol, anethole and estragol, flavonoids and other polyphenolic compounds. 10
The antiradical profile of cumin and caraway has been proposed as the underlying mechanism for their multifaceted pharmacological properties such as antimicrobial, antidiabetic, anticarcinogenic/antimutagenic, antistress, antiulcerogenic, etc. as outlined in the succeeding sections. A glucose tolerance test conducted in rabbits, cumin significantly increased the area under the glucose tolerance curve and 15 hyperglycemic peak.
A methanolic extract of cumin seeds reduced the blood glucose and inhibited glycosylated hemoglobin, creatinine, blood urea nitrogen and improved serum insulin and glycogen (liver and skeletal muscle) content in alloxan and streptozotocin (STZ) diabetic rats. 20
The collateral benefits included decreased creatinine, urea nitrogen and improved insulin and glycogen in tissue and skeletal muscles, accompanied by a reduction in rat tail tendon collagen-linked fluorescence and pepsin digestion which are implicated in the pathogenesis of diabetic microvascular complications.
Oral administration of cumin to alloxan diabetic rats reduced body weight, plasma 25 and tissue cholesterol, phospholipids, free fatty acids and triglycerides. Histological observations demonstrated significant decrease in fatty changes and inflammatory cell infiltrates in diabetic rat pancreas. Cumin suppressed alcohol and thermally oxidized oil induced hyperlipidemia.
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It decreased aspartate transaminase (AST), alkaline phosphatase (ALP) and γ-glutamyl transferase (GGT) activities and decreased the tissue (liver and kidney) levels of cholesterol, triglycerides and phospholipids and prevented the changes in the composition of fatty acids in the plasma of rats administered with alcohol and/or thermally oxidized oil. 5
Saindhav (Rock salt)
Usually, lavana rasa increases pitta dosha but due to its sheeta virya Saindhav helps to balance pitta. Due to its lavana rasa saindhav balances vata and reduces kafa. It Improves taste, digestion and strength and is good for heart.
Shunthi (Zingiber officinalis) 10
The composite methanolic extract of roots of Withania somnifera, leaves of Ocimum sanctum and rhizomes of Zingiber officinalis could be beneficial for the protection of exhaustive physical exercise like swimming induced oxidative stress injury on brain tissues and also on the vital tissues like cardiac, skeletal and hepatic which are very much prone to physical exercise-induced oxidative stress. 15
Pippali (Piper longum)
Beneficial effects of PlrAqe are especially hopeful in preventing hyperglycemia, cardiovascular, hepatic and renal diseases. In conclusion, this study has undoubtedly provided scientific confirmation and evidence for the safe use of the roots of Piper longum by traditional healers in the treatment of diabetes. 20
Marich (Piper nigrum)
Piperine has a hypoglycemic effect in normal mice. Oral administration of Piper longum dried fruits has shown significant anti-hyperglycemic, antioxidant and antilipid peroxidative effects in hyperglycemic rat’s intake of black pepper affects membrane lipid peroxidation, enzymatic and nonenzymatic antioxidants as well as 25 oxidative stress in rats.
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Treatment with P nigrum and V rosea once a day for four weeks in alloxan-induced diabetic rats effectively controls the blood glucose level as well as affects lipid level and antioxidant potential.
Ramatha (Asafoetida)
Ramatha along with few other extracts, in formulation, / chloroform, petroleum 5 ether and ethanol extracts of M. charantia Linn. and N. jatamansi has shown a significant hepatoprotective effect by decreasing the elevated serum enzyme levels such as glutamate pyruvate transaminase, glutamate oxaloacetate transaminase and alkaline phosphatase.
Table 1 below represents Properties, dosage, and pharmacological actions of 10 Ayurvedic medicines selected for the present invention.
Table 1: Properties, dosage, and pharmacological actions of Ayurvedic medicines
Name
Properties
Reported Dosage
Pharmacology
Swertia Chirayita
{chrirata ,kadechirayat}
Rasa –Tikta
Veerya –sheeta /ushna
Vipaka – katu
Guna- laghu ,
yogawahi
1 to 3 gms of powder /day is recommended.
Dry extract 78 – 260 mg in term of 1%
IP- 2-4ml.
LE- [25%Alc] 2-4 ml.
Anti-malarial property, Hypoglycemic activity,
Antiulcerogenic activity, Anti-inflammatory activity .
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Picrorhiza kurroa
{Katuka ,kutaki
Rasa- Tikta
Veerya-Sheeta
Vipaka-katu
Guna-bhedan, deepan, hrudya.
1 or 2 grams for laxative effect.
Viral hepatitis -300 mg – 500 mg 2/3 times.
SE-200-400mg
DE-150-300mg
IP-0.6-1.2ml{60% ALC}
Anti-asthmatic activity, anti-inflammatory activity, Hepatoprotective activity, Antidiabetic activity, Antibacterial activity.
Jasad bhasm (oxide of zinc)
Rasa- Katu (Spicy), Tikta (Bitter) & Kashaya (Astringent)
Virya- Sheeta (Cold)
Guna-Laghu (Light)
Vipaka- Katu
1/2-2 gunja {60-240mg}
Antacid, Anti-inflammatory
Anti-arthritic,
Anti-pyretic,
Digestive Stimulant,
Hematinic (increases hemoglobin levels),
Hematogenic (helps in formation of red blood cells)
Shankh bhasm
{clax of conch shell}
Rasa-madhur, Kashaya, katu, kshara
Guna-laghu, lekhana, khara, kshara, hima
Virya-sheet, Anushna, ushna
Vipaka –madhura, katu
1-3 gunja {120-360 mg}
Antidiarrheal, Detoxifying property, Anti-oxidant, Anti-spasmodic, Digestive stimulant, anti-emetic, stool bind agent.
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Zingiber officinale
{ginger/suntha}
Rasa-katu,
Veerya- ushna,
Vipaka-madhur,
Guna- Deepan,bhedan,ushna,vaatkapha shamak
DS -0.3-10.0 gms
SE 1500-4800 in term of 1% extractives
For allergy/ motion sickness -9000mg of 1%
Strong Tr 0.3-0.6ml
Oleoresin 16-60 mg.
Anti-emetic activity, Anti-inflammatory activity, Anti-coagulant, cardiovascular effects,
Antitumor property, Antimicrobial activity, Miscellaneous activity.
Piper longum
{Indian long pepper,pimpali}
Rasa-katu,
Veerya- Anushna and sheet when fresh,
Vipaka – madhur,
Guna-laghu,snegdha,tikshna
Powder of piper longum 0.3-0.6 gm/day can be recommended
SE-45-90mg,
Decoction{1/10}IPC-50-60 ml.
Anti asthamatic activity,Cardio vascular activity, Effect on central nervous system,Anti fertility activity, Anti-bacterial activity ,Anti tubercular activity ,Anti-inflammatory effects.
Piper Nigrum
{ black pepper,mire}
Rasa-kattu,
Veerya –ushna ,
Vipaka –katu,
Guna-laghu,tikshna ,rooksha,deepan
0.3 to 0.6 gm powder with honey is recommended /day
SE-30-60mg,confection of pepperIPC-4-8 gm.
Anti-convulsant activity, Anti-malarial activity, Anti-microbial activity.
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Allium sativum
{garlic ,lasun}
Rasa-pancha rasa except lavana
Veerya-ushna
Vipaka- katu
Guna-ushna,tikshna
Typical dosage - 900 mg/daily.
Anti-astherosclerosis activity, Antithrombotic, Hypoglycemic ,anti oxidant, Anti-cancer, Anti-microbial, Anti tuberculosis effect.
Cuminum cyimum
{cumin seed, jeera}
Rasa- katu ,
Vipaka –katu
Veerya –ushna ,
Guna –laghu ,rooksha
1-2 masha
{3-6 gms one or two times}
Appetizer,carminative,blood purifier, Anti spasmodic , Anti fungal.
Asafoetida ferula {hing}
Rasa- katu,
Veerya –ushna
Vipak – katu
Guna –laghu, snigdha , Teekshna ,
2-8 ratti {240mg-960mg}
Antibacterial, Antifungal, Antimicrobial ,hypotensive ,Anticancer,Anti diabetic , Antispasmodic.
Sendha namak
{halite}
Rasa –lavan
Veerya –sheet
Reduced risk of sodium level, Anti inflammatory ,respiratory disorders.
The above Ayurvedic medicines are reportedly effective in large doses such as preferably 300 mg and above, more preferably 500 mg and above and most preferably 1 g and above.
For example, recommended dose of Swertia Chirayita 1-3 g per day, of Picrorhiza 5 kurroa is 1-2 g / day and Cumin is from 3 – 6 g one or two times in a day. It is not
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possible to combine these medicines in such higher amounts to produce a formulation which can be conveniently administered.
It is further highly desirable that the formulation is patient compliant and shall be consumed by the patients with ease for days or months. Therefore, the size of formulation dose should be such that it can be effectively consumed. If each 5 ingredient is required in an amount of from 0.5 g – 5 g, such formulation would be bulky and would not have patient compliance.
Additionally, Ayurvedic formulation may contain one or more inactive ingredients such as lactose, starch, gum, talc, stearates, colloidal silicon dioxide, sugar, sugar alcohol, saccharide, cellulose, cellulose based binder and / or disintegrants, fibre, 10 one or more polymers etc.
Additionally, or alternatively, Ayurvedic formulation may contain one or more inactive ingredient / Ayurvedic excipients such as Guggulu, Honey, Turmeric, Ghee, takra, jala, cow urine, pippli, Adrakswarasa, Mulethi, Dalchni, Dhanyak, Trikatu, Amla juice etc. 15
One of the preferred formulation is an Ayurvedic capsule. Another preferred formulation is a tablet. Yet another preferred formulation is powder packed in a Sachet.
Alternatively, liquid formulations such as syrup, suspensions or any other oral delivery form can be employed. 20
Amount of inactive ingredients is usually from 0-25 %, preferably, from 0-20 % and more preferably, from 0-15 % of the compositions. Here, composition means a composition which is ready to be filled in the capsule or ready to be compressed into a tablet or etc.
The inventors have found that capsules can be manufactured using from 0-10 % 25 inactive ingredients and tablets can be manufactured using from 0-15 % inactive ingredients of the composition ready for encapsulation or tableting.
Ayurvedic formulation of the present invention accommodates 11 Ayurvedic medicines in a single composition or 2-4 different compositions.
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Each composition can be a powdered or granular composition or tablets / pills. The powdered composition(s) can be filled in sachet or encapsulated in a hard gelatin or vegan / vegetarian capsule like HPMC capsule.
The preferred formulation is a capsule containing single or 2-4 compositions preferably in powder form. 5
In an embodiment, the Ayurvedic formulation has all 11 Ayurvedic medicines in a capsule of size 00 or 000. This embodiment is presented under example 1, table 2.
In another embodiment, the Ayurvedic formulation has two compositions, each having 3 – 8 ingredients wherein each composition is in a capsule of sizes 0 or 1. This embodiment is presented under example 2, table 3. 10
In yet another embodiment, the Ayurvedic formulation has three compositions, each having 3 – 5 ingredients wherein each composition is filled in a capsule of sizes 1 or 2. When the weight to be filled is in the range of 150 – 250 mg, size 3 capsule also can be employed. This embodiment is presented under example 3, table 4A. 15
In one more embodiment, the Ayurvedic formulation has four compositions, each having 2 – 5 ingredients wherein each composition is filled in a capsule of sizes 3. When the weight to be filled is in the range of 150 – 250 mg, size 3 capsule also can be employed. This embodiment is presented under example 3, table 4B.
The capsule size depends on density of enclosed composition which can vary 20 between 0.25 g / ml – 1.2 g/ml.
A single formulation having all 11 Ayurvedic medicines including herbal and mineral medicines accommodated in a single capsule of size 00 when the weight of capsule content is up to 1000 mg or size 00 when the weight of capsule content is greater than 1000 mg and up to 1500 mg. 25
Types of capsule formulations are provided under examples 1-3. They will be manufactured on large scale, packed, labelled as per the GMP and GCP regulations and requirements. Storage will be at a controlled temperature and moisture and away from direct Sunlight and heat.
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When Ayurvedic formulation of the present invention which is in the form of a single composition or 2-4 separate compositions is administered for minimum 3 days but preferably at least 7 days, preferably for at least 15 days, more preferably for at least 30 days, most preferably for at least 60 - 90 days, it relieves patients 5 from elevated levels of one or more of LDL, triglycerides, blood glucose, HbA1c, bilirubin (total and direct), alanine transaminase (ALT) and aspartate transaminase (AST), alkaline phosphatase (ALP).
Formulation 2 of example 2 having two separate compositions as IP1 and IP2 was taken for a randomized, double-blind, parallel-group study to determine the efficacy 10 and safety of polyherbal formulation in patients suffering from NAFLD. The demographic and anthropometric parameters of patients selected are presented in table 5.
In the double-blind study, the formulation 2 i.e. two separate capsules having composition 1 and composition 2 are administered twice in a day to patients 15 suffering from NAFLD for a period of 90 days. The study is provided under example 5.
Patients Inclusion Criteria for the double blind study:
i) target patients’ size - 82 patients;
ii) Age - Patients were in the age range of 18-70 years. 20
iii) Patients with diabetes and hyperlipidemia as comorbidities;
iv) patients previously diagnosed with fatty liver based on
a) > 20% liver fat on MRI-PDFF or liver fat > 33% on biopsy within the past six months;
b) If MRI-PDFF at the screening was between 15-18%, then those 25 patients have ALT levels > 30 U/L for males and > 19 U/L for females;
c) BMI between 25-40 kg/m2, and
d) HbA1c between 5.7 – 7%
were considered for the study 30
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The demographic and anthropometric parameters of patients selected are presented in table 5.
Exclusion criteria include patients with a known history of hypersensitivity to herbal extracts, jaundice for more than 3 months, advanced fibrosis, cirrhosis, and 5 other liver diseases. Pregnant or lactating females, patients on immuno-modulatory drugs, severe renal, hematopoietic disease or cardiac insufficiency, any form of complementary alternative medicine, and those undergone liver transplants.
Administration of Polyherbal / Ayurvedic formulation 10
The patients were randomized to two arms using block randomization of size two. These two arms are
First arm - IP+SOC; and Second arm - Placebo +SOC.
SOC is standard of care treatment and comprises of administering medicines, Metformin, Glycomet, Telmikind, Stamlo, Vit E 400mg, Atorva 10mg Lipaglyn 4 15 mg to both the arms. This means standard medicines were received by all patients. In addition, the patients in the IP+SOC arm received 1800 mg of ayurvedic formulation of example 2, with a dosing pattern of 450 mg of IP1 capsule + 450 mg of IP2 capsule in the morning and the same in the evening.
The patients in the placebo arm received dosing of placebo contents of 1800 mg of 20 example 4 along with SOC. The contents of investigational products and placebo are given under example 2, and example 4 respectively. The patients in each arm were advised to take the doses every day for 90 days along with SOC. The blood samples not exceeding 30 ml were collected at baseline and day 90 for evaluating the efficacy and safety of parameters. All the patients were asked to visit the site on 25 days 30, 60, and 90 for assessments. The reduction in all these parameters is statistically significant with P-value < 0.05.
To assess the efficacy of The Ayurvedic formulation, the target primary endpoints were: total bilirubin, direct bilirubin, alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), anti-oxidant enzymes 30
20
like malondialdehyde (MDA), superoxide dismutase (SOD), Catalase, Glutathione (GSH) and Glutathione peroxidase.
Additionally, hematological parameters like Hb, PCV, WBC-DLC, MCV, MCH, MCHC, and MPV were also assessed. The lipid profile including LDL, HDL, TG, and TC as well as blood chemistry parameters like fasting glucose and HbA1c were 5 also evaluated.
The secondary endpoints involved MRI and ultrasound grading (USG). The USG grading was based on diffuse hepatic steatosis and was categorized into grades I – III.
Grade I – III fatty liver are described below: 10
Grade I - This is the mildest form. Here, the fat accumulates on the outside of the organ and does not affect its function.
Grade II - It is a moderately severe form and requires medical intervention to prevent worsening.
Grade III - It is the severe form of fatty liver, with the appearance of symptoms. 15
All the primary and secondary endpoints were evaluated at baseline (after randomization) and end-of-study. Safety assessment included vital parameters, physical examination and any other adverse event (AE) reported at the time of clinical examination. 20
The subjects were followed till the end of the study period. Figure 2 shows the study flow diagram. The entire study was conducted following the principles of Good Clinical Practice (GCP) and the current version of the Declaration of Helsinki, ICMR (Indian Council of Medical Research) Ethical Guidelines for Biomedical Research on Human patients (2017), and applicable regulatory guidelines. The data 25 generated on demographic, anthropometric, and various laboratory parameters were quality-checked and subjected to statistical analysis.
Statistical analysis
The continuous parameters were expressed in terms of mean, standard deviation, 30 and 95% confidence interval, while categorical parameters were summarized as
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frequencies and percentages. The between-group comparison of continuous parameters was carried out using a t-test for independent samples or the Wilcoxon rank sum test, based on the assumption of normality. For between-group comparisons, the values of continuous variables at day 90 were adjusted with screening values using a one-way analysis of covariance (ANCOVA). The 5 comparison of raw values of the parameter between screening and day 90 was performed using paired t-test.
Tests conducted at baseline and at 90 days include
i) liver function tests typically including alanine transaminase (ALT) and aspartate transaminase (AST), alkaline phosphatase (ALP), serum 10 bilirubin and total bilirubin.
ii) Levels of anti-oxidant enzymes;
iii) haematological parameters;
iv) lipid and glucose parameters;
It has been surprisingly found that the Ayurvedic formulation of the present 15 invention provides following results within the group:
a) Reduction in each of alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), total bilirubin and direct bilirubin;
b) Enhancement in levels of each of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) (anti-oxidant enzymes). 20
c) Reduction in total cholesterol, LDL cholesterol and HbA1c.
The enhancement in the levels of anti-oxidant enzymes indicate readiness for the catalytic removal of free radicals and reactive species by the body. This enhancement in the levels of anti-oxidants is statistically significant with P-value < 0.05. 25
Between the groups, reduction in direct bilirubin was significant.
The results of all above studies are reported in tabular form from tables 6 – 9.
Additionally, results of MRI studies and ultrasound grading (USG) are provided under figures 3 and 4 respectively.
22
Results of a randomized, double-blind, parallel-group study to determine the efficacy and safety of polyherbal formulation in patients suffering from NAFLD.
Eighty-two patients were randomized equally in two groups. In the SOC+IP group, 5 (first arm of the study) liver function parameters showed statistically significant improvement (p < 0.05) from baseline to end-of-study, unlike the SOC+Placebo group (second arm of the study). The anti-oxidant enzyme catalase showed significant improvement in the SOC+IP group (p = 0.01). Further, TC, LDL, and HbA1c improved significantly in the SOC+IP group (p < 0.05). The proportion of 10 patients showing improvement in the grade of fatty liver in the SOC+IP group was significantly higher than the SOC + Placebo group (p = 0.003).
Results
15
The analysis set consisted of 82 patients (41 in each arm) without any loss to follow-up. The mean age of the patient in the SOC+IP arm was 43.17 ± 8.99 years, while that of the SOC + Placebo arm was 41.83 ± 11.12 years, and the difference in the means was statistically non-significant (Table 5). The mean BMI in the SOC+IP arm was 28.51 ± 2.87 kg/m2, while in the SOC + Placebo arm was 29.35 ± 4.05 20 kg/m2, and the difference was statistically non-significant. The difference in gender distribution between the two arms was also statistically non-significant. Thus, the baseline demographic and anthropometric parameters were similar in the two arms.
Table 6 gives the between and within-group comparison of liver function test (LFT) parameters. The between-group comparison was performed at both baseline and 25 day 90. At baseline, all the LFT parameters showed statistically non-significant differences between the two arms, except enzyme catalase (p = 0.013). On day 90, the significance values for each parameter were obtained after adjustment with respective baseline values. All the LFT parameters and anti-oxidant enzymes showed statistically non-significant differences between the two arms (p > 0.05), 30 except direct bilirubin . The adjusted mean for direct bilirubin in SOC+IP arm was
23
significantly smaller than the SOC + Placebo arm (p = 0.023). The within-group comparison of parameters in the SOC+IP arm revealed a statistically significant reduction in ALP, ALT AST, Total and direct bilirubin parameters at day 90 from baseline (p < 0.05). However, the change of these parameters in the SOC + Placebo arm was statistically non-significant. The anti-oxidant enzymes MDA and SOD 5 showed statistically significant change from baseline to day 90 in both arms (p < 0.05). The parameter catalase showed significant improvement in the SOC+IP arm (p = 0.01), while the change was statistically non-significant in the SOC + Placebo arm.
The comparison of hematological parameters between groups revealed statistical 10 non-significance at day 90, after adjustment with respective baseline values (Table 8). The within-group comparison resulted in a significant reduction in WBC-DLC (p = 0.038) in the SOC+IP arm ; although, the mean was in the clinically acceptable range. The other parameters showed statistically non-significant change between two-time points in both arms. 15
The lipid parameters were also compared between and within the group as shown in Table 9. The difference of means between the two arms for all the parameters at day 90, after adjusting with the respective baseline, was statistically non-significant. The within-group comparison revealed a statistically significant reduction in Total cholesterol, LDL, and HbA1C from baseline to day 90 in the SOC+IP arm with p-20 values < 0.05. The change for these parameters in the SOC + Placebo arm was non-significant.
Figure 3 gives the bar chart representation of the mean of MRI finding at two-time points in the two arms. In the SOC+IP arm, there was a statistically significant change in MRI findings with a median of 22 at baseline to a median of 19 at day 25 90, with a p-value of 0.007. In the SOC + Placebo arm, there was a change from a median of 21 at baseline to 18.5 at day 90, with a p-value of 0 .018. Thus, in both arms, there was a significant change from screening to day 90. The difference in medians between the two arms at both baseline and day 90 was statistically non-significant. The comparison of USG-based grades was performed between two 30
24
arms on day 90, as shown graphically in Figure 4. All 41 patients in both arms had grade II at baseline. On day 90, in the SOC+IP arm, 23 (56.1%) had grade I, while in the SOC + Placebo arm, 10 (24.4%) had grade I. This is an excellent finding that severity of fatty liver diseases has reduced. The difference in proportions between the two arms was statistically significant with a p-value of 0.003. 5
As regards safety, 21 adverse events in 18 patients were reported in the study. In the SOC+IP arm, there were 6 AEs, while in the SOC + Placebo arm, there were 11 AEs. The random blood glucose showed reduction in mean glucose level at day 90 in both the groups, however the change was statistically non-significant. All the adverse events were mostly general disorders like headache, giddiness & cough, 10 and were in mild form.
Discussion
NAFLD is a reversible condition in which large vacuoles of triglyceride fat accumulate in the liver through a process called steatosis, due to causes other than 15 alcohol use. It is a component of metabolic syndrome comprising central obesity, hypertension, impaired fasting glucose, impaired glucose tolerance, hypertriglyceridemia, and low HDL (Cankurtaran M et al, 2007). The manifestation is mainly due to excessive caloric intake, and lack of physical activity thereby requiring correction in the lifestyle, as the first level of prevention and treatment. 20 However, when lifestyle changes are insufficient, external medication support becomes the strategic line (Centis E et al, 2010). Fatty liver needs early treatment to avoid terminal liver disease. The SOC treatment majorly comprises Metformin, Glycomet, Telmikind, Stamlo, Vit E 400mg, Atorva 10mg and Lipaglyn 4 mg, to control blood sugar levels, hypertension and treat non-cirrhotic non-alcoholic 25 steatohepatitis. The etiology, pathogenesis, and symptomatology of NAFLD does not appear as a single entity, but from the Ayurveda perspective, it can be described under broad categories of Agnivikriti (deranged digestion) and Medoroga (disturbed fat metabolism) (E. Ramya et al, 2021). The complex pathogenesis of the disease involves oxidative stress, inflammation, lipotoxicity, and insulin 30
25
resistance (Buzzetti E. et al, 2016). Hence, a combination of agents is required that can act through different mechanisms to produce the desired effect. In the present study, the investigational product is a combination of medicinal herbs, seeds, roots, and dried fruits. One of the main ingredients of formulation IP1 is the herb Swertia Chirayita (Gentianaceae). It has demonstrated a good anti-inflammatory activity 5 (Verma H.et al, 2008). The anti-hepatitis B virus activity of S. chirayita extracts was also studied on HepG 2.2.15 cells line (Zhou N. J., 2015). Another medicinal plant Picrorhiza kurroa regressed several features of NAFLD like lipid content of the liver tissue, morphological regression of fatty infiltration, hypolipidemic activity, and cholestatis . It has properties of tikta rasa (bitter taste), lagu ruksha 10 guna (lightness quality), and katu vipaka (absorption of water and electrolyte in large intestine), which supports its pharmacodynamic activity on lipids in case of lipid disorders (Koolman AH., 2010).
In an animal-based study of standardized extracts of P kurroa in experimental NAFLD, the authors observed a reduction in alkaline phosphatase (ALP) to normal 15 levels in the P. Kurroa 200 mg/kg and 400 mg/kg groups, unlike in the other experimental groups (Shetty S. et al, 2010).
The authors also observed a significant reduction in ALT levels at the end of 6 weeks, as compared to baseline (p < 0.001). Another herb Ramatha (family name: Ferula narthex) has the hepato-protective effect that decreases the elevated serum 20 enzyme levels such as glutamate pyruvate transaminase, glutamate oxaloacetate transaminase, and alkaline phosphatase (Nabi SA et al, 2013).
The present study also showed a significant reduction in mean ALP and ALT levels in the SOC+IP arm from baseline to day 90, which has P. Kurroa as well as Ramatha as ingredients. Moreover, in the SOC+IP arm, there was a significant reduction in 25 the AST levels at day 90 as compared to baseline, which was not observed in the SOC + Placebo arm. The effect of SOC+IP on bilirubin was also statistically significant.
Cuminum cyminum (Jiraka), showed significant antioxidant activity (Srinivasan K et al, 2005). This is one of the ingredients of IP2. There was a significant increase 30
26
in the mean oxidative stress marker of malondialdehyde (MDA) in the SOC+IP arm, which could be attributed to cumin.
In an animal study conducted by Miah P. et al. (2021), the authors observed a significant increase in the MDA levels with cumin powder supplementation at day 90 as compared to baseline. Moreover, the enzymes Superoxide dismutase (SOD) 5 and catalase showed a significant increase in both plasma and liver after administering cumin in high-fat diet-fed rats. We also observed significantly increased levels of catalase and SOD at day 90 from baseline in the SOC+IP arm. Catalase is a key antioxidant enzymes that mitigates the oxidative stress by decomposing cellular H2O2. The results for MDA and SOD were also significant 10 in the SOC + Placebo arm, except for catalase. The mean glutathione (GSH) levels were nearly unaltered at day 90 in both the treatment arms; however, in the above study, the authors observed depletion in high-fat diet rats treated with cumin as compared to controls. Allium sativum (Garlic), which is also one of the key ingredients of IP2, has a promising lipid-lowering effect on hyperlipidemia patients 15 (Memon A.R. et al, 2018).
In the study by Soleimani D et al. (2020), the authors assessed the efficacy of garlic on hepatic steatosis in patients with NAFLD. After adjusting with energy intake and physical activity, the metabolic factors, except HDL, showed significant change from baseline to week 15 in the garlic treated group. 20
In a study by Kwak JS et al. (2014), the garlic powder intake significantly reduced LDL cholesterol by 15.8 mg/dl and total cholesterol by 8.1 mg/dl, without changing serum HDL cholesterol.
In present study also, the inventors observed a reduction in total cholesterol (22.11 mg/dl) and LDL (14.75 mg/dl) by day 90 in the SOC+IP arm, which could be due 25 to the presence of garlic in IP2 . Garlic decreases the absorption and synthesis of cholesterol, thereby decreasing the levels of cholesterol.
The human enzymes required in the cholesterol biosynthesis, such as squalene monooxygenase and HMG-CoA reductase are inhibited by the constituents of
27
garlic. However, non-significant changes were seen in SOC+Placebo arm. The presence of Zingiber officinalis (Shunthi) extract in IP2 helps in improving endurance capacity by minimizing oxidative stress on vital organs (Lemhadri A. et al, 2006). Earlier animal studies have shown that ginger enhances antioxidant defense system such as glutathione peroxidase and glutathione S-transferase 5 (Motwani TK et al, 2011). Moreover, ginger inhibits pathways in the inflammatory process i.e. arachidonic acid metabolism, thus exhibiting anti-inflammatory properties (Shahid A et al, 2020). The authors observed significant improvement in the fatty liver grading (56.7%) in the ginger ingredient-based test group (Murabba-I Zanjabil) as compared to the control group, on the ultrasonogram. 10
In present study, in the SOC+IP arm, there was a significant improvement in the grade of NAFLD on USG from II to I by day 90, as observed through ultrasonography (56.1%), unlike the SOC + Placebo arm showing improvement in 24.4% of patients . The effect could be attributed to Zingiber officinalis in IP2. Another ingredient of the product Piper longum (Pippali) has been effective in 15 preventing hyperglycemia, cardiovascular, hepatic, and renal diseases . Piper nigrum (black pepper) also has shown anti-hyperglycemic, anti-oxidant, and anti-lipid peroxide effects. It exerts action by stimulating the function or number of beta cells and thus increasing the insulin secretion with least side effects. We observed a significant reduction in HbA1c in the SOC+IP arm, which could be attributed to 20 the above two ingredients . Moreover, IP1 contains calx of conch shell, which has strong antacid and anti-oxidant properties.
Regarding safety, all the reported adverse events were general disorders and in a mild form. There was no drug-drug (allopathy and herbal) interaction observed during the study. 25
One limitation of the study was that MRI, which is the gold standard, could be performed only on 26 patients from both arms; hence, statistical significance of difference in the findings was missing. More clinical trials with robust designs and larger sample sizes are needed in which lifestyle modification aspects also should be addressed. 30
28
Conclusion: The etiology and pathogenesis of NAFLD cannot be treated with a single entity, but requires a combination of multiple ingredients. The group exposed to polyherbal formulation showed statistically significant improvement in the liver, lipid parameters, and grades as compared to the Placebo group, and hence we believe that the formulation can be recommended for treating NAFLD in addition 5 to standard care. Following examples illustrate the invention without restricting it’s scope.
Example 1 - Ayurvedic formulation having a single composition
Table 2: Formulation containing Single composition having 11 Ayurvedic 10 medicines
Formulation / Sr. No.
Ingredient
Latin Name
Preferred range
Quantity in mg
Formulation
Single composition
Kirat- tikta
(panchang)
Swertia chirata
150 – 250 mg
210
(23.6 %)
Katuka
(root)
Picrorrhiza kurroa
125 – 225 mg
150
(16.85 %)
Jasad bhasm
(Oxide of zinc)
10 – 35 mg
30
3.37 %
Shankh bhasm
(Calx of conch shell)
20 – 70 mg
60
(6.74 %)
Lashun
Alium sativum
100 - 200 mg
140
(15.73 %)
Jiraka
Cuminum cyminum
20 - 80 mg
70
(7.86 %)
Saindhav
(Rock salt)
20 - 80 mg
30
(3.37 %)
29
Shunthi
Zingiber officinalis
20 - 80 mg
70
(7.86 %)
Pippali
Piper longum
20 - 80 mg
30
(3.37 %)
Marich
Piper nigrum
20 - 80 mg
70
(7.86 %)
Ramatha
Asafoetida
20 - 80 mg
30
(3.37 %)
Total weight of the composition
890
Example 2 - Ayurvedic formulation having two compositions
Table 3: Formulation containing two compositions having a total of 11 Ayurvedic medicines 5
Formulation / Sr. No.
Ingredient
Latin Name
Quantity
Quantity in mg
Formulation having 2 compositions
Composition 1 /IP1
Kirat- tikta
(panchang)
Swertia chirata
150 – 250 mg
200
(22.22 %)
Katuka
(root)
Picrorrhiza kurroa
125 – 225 mg
180
(20 %)
Jasad bhasm
(Oxide of zinc)
10 – 35 mg
20
(2.22 %)
Shankh bhasm
(Calx of conch shell)
20 – 70 mg
50
(5.55 %)
30
Formulation / Sr. No.
Ingredient
Latin Name
Quantity
Quantity in mg
Composition 2 /IP2
Lashun
Alium sativum
100 - 200 mg
150
(16.66 %)
Jiraka
Cuminum cyminum
20 - 80 mg
50
(5.55 %)
Saindhav
(Rock salt)
20 - 80 mg
50
(5.55 %)
Shunthi
Zingiber officinalis
20 - 80 mg
50
(5.55 %)
Pippali
Piper longum
20 - 80 mg
50
(5.55 %)
Marich
Piper nigrum
20 - 80 mg
50
(5.55 %)
Ramatha
Asafoetida
20 - 80 mg
50
(5.55 %)
Total weight of the two compositions
900 (100 %)
Example 3 - Ayurvedic formulation having 3 - 4 compositions
Table 4A: Formulation containing three compositions having a total of 11 Ayurvedic medicines 5
Formulation / Sr. No.
Ingredient
Latin Name
Preferred range
Quantity in mg
Formulation having 3 compositions
31
Composition 1 /IP1
Kirat- tikta
(panchang)
Swertia chirata
150 – 250 mg
180
(19.78 %)
Katuka
(root)
Picrorrhiza kurroa
150 – 225 mg
200
(21.98 %)
Jasad bhasm
(Oxide of zinc)
10 – 35 mg
10
(1.1 %)
Shankh bhasm
(Calx of conch shell)
20 – 70 mg
40
(4.40 %)
Composition 2 /IP2
Lashun
Alium sativum
100 - 200 mg
180
(19.78 %)
Jiraka
Cuminum cyminum
20 - 80 mg
30
(3.30 %)
Saindhav
(Rock salt)
20 - 80 mg
70
(7.69 %)
Composition 3 /IP3
Shunthi
Zingiber officinalis
20 - 80 mg
30
(3.30 %)
Pippali
Piper longum
20 - 80 mg
70
(7.69 %)
Marich
Piper nigrum
20 - 80 mg
30
(3.30 %)
Ramatha
Asafoetida
20 - 80 mg
70
(7.69 %)
Total weight of 3 compositions
910 (100 %)
Example 3 - Ayurvedic formulation having 3 - 4 compositions
Table 4B: Formulation containing four compositions having a total of 11 Ayurvedic medicines 5
32
Formulation / Sr. No.
Ingredient
Latin Name
Preferred range
Quantity in mg
Formulation having 3 - 4 compositions
Composition 1 /IP1
Kirat- tikta
(panchang)
Swertia chirata
150 – 250 mg
180
(19.78 %)
Shankh bhasm
(Calx of conch shell)
20 – 70 mg
30
(4.40 %)
Composition 2 /IP2
Jasad bhasm
(Oxide of zinc)
10 – 35 mg
10
(1.1 %)
Katuka
(root)
Picrorrhiza kurroa
150 – 225 mg
200
(21.98 %)
Composition 3 /IP3
Lashun
Alium sativum
100 - 200 mg
180
(19.78 %)
Jiraka
Cuminum cyminum
20 - 80 mg
30
(3.30 %)
Saindhav
(Rock salt)
20 - 80 mg
70
(7.69 %)
Composition 4 /IP4
Shunthi
Zingiber officinalis
20 - 80 mg
30
(3.30 %)
Pippali
Piper longum
20 - 80 mg
70
(7.69 %)
Marich
Piper nigrum
20 - 80 mg
30
(3.30 %)
Ramatha
Asafoetida
20 - 80 mg
70
(7.69 %)
Total weight of 3 compositions
900 (100 %)
33
Process of preparing compositions of examples 1-3.
1) Coarse form of herbal medicines are dispensed separately according to their weights in a container;
2) Bhasma form and fine rock salt are dispensed separately.
3) Each coarsely powdered medicine is subjected to grinding in a grinder to 5 produce finely ground microfine form;
4) Microfine powdered medicine is subjected to hydroalcoholic extraction using cold percolation method;
5) Hydroalcoholic extract of medicine is subjected to hydro distillation in a hydro-distillation machine to obtain dry extract containing little or no 10 solvent;
6) If required, extracts are further dried to produce dry powder;
7) Dried extracts, Bhasma form of and powdered form of rock salt are ground again in a grinder, sifted and transferred to a blender based on the final composition / compositions to blend and produce uniform blend of 15 compositions of example 1or 2 or 3 containing all 11 ingredients in a single composition or 2-4 different compositions; and
8) Optionally the blend / blends of step 7 is / are blended with one or more inactive ingredients;
9) The blend is filled into a sachet, compressed into a tablet or filled into a 20 capsule. Capsule is a hard gelatin or cellulose based Capsule.
Example 4 – Placebo Capsules
Process
1. Desired quantities of Lactose monohydrate, maize starch and 25 croscarmellose sodium are weighed and blended in a blender;
2. Around 900 mg of this placebo blend is filled in 1, 2 or 3 capsules to prepare placebo capsules.
Example 5 30
34
Formulation 2 of example 2 having two separate compositions as IP1 and IP2 was taken for a randomized, double-blind, parallel-group study to determine the efficacy and safety of Ayurvedic formulation / poly-herbomineral formulation in patients suffering from NAFLD.
5
Table 5 represents demographic and anthropometric parameters at baseline.
Example 6
Table 6 : Between and within group comparison of liver parameters 10
35
Table 7 - Between and within group comparison of anti-oxidant enzymes
36
37
Table 8 : Between and within group comparison of haematological parameters
*Obtained using paired t-test; †Obtained using t-test for independent samples after adjusting with respective baseline values of parameter using ANCOVA; Bold p-values indicate statistical significance 5
38
Table 9 : Between and within group comparison of lipid and glucose parameters
P-value below 0.05 is statistically significant.
5
39
Table 10 : Description of investigational products
Investigational products
Product
IP+SOC
Placebo+SOC
Name
IP1 + IP2 + Standard of Care Treatment
Placebo 1 + Placebo 2 Standard of Care Treatment
Composition
a. IP1
Composition 1 of example 2
b. IP2
Composition 2 of example 2
Composition of example 4
Each Placebo Capsule is ~ 450 mg.
Dosage form
Capsule
Capsule
Dosage
Two capsules X 2 times a day
1800 mg/ day along with SOC
Two placebo capsules X 2 times a day
1800 mg/ day along with SOC
Administration
Oral administration
Oral administration
SOC includes administering following medicines, Metformin, Glycomet, Telmikind, Stamlo, Vit E 400mg, Atorva 10mg Lipaglyn 4 mg.
40
References
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Supplementation of cumin seed power prevents oxidative stress, hyperlipidemia and non-alcoholic fatty liver in high fat diet fed rats. Biomedicine & Pharmacotherapy. 2021: 141. 5
Karan M, Vasisht K, Handa SS. Antihepatotoxic activity of Swertia chirata on carbon tetrachloride induced hepatotoxicity in rats. Phytother Res. 1999;13(1):24-30. doi:10.1002/(SICI)1099-1573(199902)13:1<24::AID-PTR378>3.0.CO;2-L
Kaushal, K., H. Singh, and A. K. Thakur. “ANTIOXIDANT AND HEPATOPROTECTIVE EFFECT OF SWERTIA CHIRATA ON HYPOXIA-10 INDUCED OXIDATIVE STRESS IN WISTAR RATS”. Asian Journal of Pharmaceutical and Clinical Research, vol. 12, no. 1, Jan. 2019, pp. 76-80, doi:10.22159/ajpcr.2019.v12i1.28502.
Sanaz Pourreza, Pouria Sefidmooye Azar, Sarvin Sanaie, Nooshin Noshadi, Saeedeh Jalali, Hamid Reza Niazkar, Arash karimi, Mahdi Vajdi, "Therapeutic 15 Effects and Mechanisms of Action of Garlic (Allium sativum) on Nonalcoholic Fatty Liver Disease: A Comprehensive Systematic Literature Review", Evidence-Based Complementary and Alternative Medicine, vol. 2022, Article ID 6960211, 15 pages, 2022. https://doi.org/10.1155/2022/6960211.
Soleimani D, Paknahad Z and Rouhani MH. Therapeutic effects of garlic on hepatic 20
steatosis in non-alcoholic fatty liver disease patients: A randomized clinical trial.
Diabetes Metab Syndr Obes. 2020; 13: 2389-2397.
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factors: a meta-analysis of randomized controlled clinical trials. Nutr Res Pract.
2014;8(6):644–654. 25
Cankurtaran M, Tayfur O, Yavuz BB, Gevik S, Akhan O, Arslan S. Insulin resistance and metabolic syndrome in patients with NAFLD but without diabetes: effect of a 6 month regime intervention. Acta Gastroenterol Belg. 2007;70:253–9.
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lifestylechanges in non-alcoholic fatty liver disease. Dig Dis 2010;28:267-73.
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formulation) and lifestyle modification in the management of non-alcoholic fatty liver disease – A randomized placebo-controlled clinical trial. AYU. 2021; 41:98-5 106.
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Verma H., Patil P. R., Kolhapure R. M., Gopalkrishna V. (2008). Antiviral activity of the Indian medicinal plant extract, Swertia chirata against herpes simplex 10 viruses: a study by in-vitro and molecular approach. Indian J. Med. Microbiol. 26, 322–326.
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(2015). Anti- hepatitis B virus active constituents from Swertia chirayita. Fitoterapia 100, 27–34. 15
Koolman AH, Bloks VW, Oosterveer MH, Jonas I, Kuipers F, Sauer PJ, et al.
Metabolic responses to long-term pharmacological inhibition of CB1-receptor activity in mice in relation to dietary fat composition. Int J Obes (Lond) 2010;34:374–84.
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Picrorhiza Kurroh Rolay ex Benth in experimental non-alcoholic fatty liver disease. J Ayurveda and Integr Med. 2010: 1(3); 203-210.
Nabi SA, Kasetti RB, Sirasanagandla S, Tilak TK, Kumar MV, Rao CA. Antidiabetic and antihyperlipidemic activity of Piper longum root aqueous extract in STZ induced diabetic rats. BMC Complement Altern Med. 2013;13:37. 25
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Shahid A, Riyazuddin M, Siddiqui MA. Effect of Zanjabil (Zingiber officinale) in
non-alcoholic fatty-liver disease- A randomized controlled trial. 2020; 8(2): 43-53.Claims We claim
1. An ayurvedic formulation characterized in having a total of eleven ayurvedic herbal and / or mineral medicines contained in 1 – 4 compositions for management of Non-alcoholic fatty liver diseases.
2. The ayurvedic formulation as claimed in claim 1 when present as 2 – 4 separate compositions, each composition has 2 – 8 medicinal ingredients.
3. The ayurvedic formulation as claimed in claim 1 wherein eleven ayurvedic herbal and / or mineral medicines include Kirat- tikta (Swertia chirata), Katuka (Picrorrhiza kurroa), Lashun (Alium sativum), Jiraka (Cuminum cyminum), Shunthi (Zingiber officinalis), Pippali (Piper longum), Marich (Piper nigrum), Ramatha (Asafoetida), Jasad bhasm (Zinc oxide) and Shankh bhasm (Incinerated Conch shell).
4. The ayurvedic formulation as claimed in claim 1 wherein herbal medicines include Kirat- tikta (Swertia chirata), Katuka (Picrorrhiza kurroa), Lashun (Alium sativum), Jiraka (Cuminum cyminum), Shunthi (Zingiber officinalis), Pippali (Piper longum), Marich (Piper nigrum), Ramatha (Asafoetida). The mineral medicines include Jasad bhasm (Zinc oxide) and Shankh bhasm (Incinerated Conch shell).
5. The ayurvedic formulation as claimed in claim 1 wherein mineral medicines include Jasad bhasm (Zinc oxide) and Shankh bhasm (Incinerated Conch shell).
6. The ayurvedic formulation as claimed in claim 1 having a total of eleven ayurvedic herbal and / or mineral medicines contained in a single composition wherein the single composition is selected from a powder composition, a tablet / pill composition, or a capsule composition or a syrup composition.

7. The ayurvedic formulation as claimed in claim 1 having a total of eleven ayurvedic herbal and / or mineral medicines contained in two separate compositions wherein each of the two compositions is selected from a powder composition, a tablet / pill composition, or a capsule composition or a syrup composition.
8. The ayurvedic formulation as claimed in claim 1 having a total of eleven ayurvedic herbal and / or mineral medicines contained in three separate compositions wherein each of the three compositions is selected from a powder composition, a tablet / pill composition, or a capsule composition or syrup composition.
9. The ayurvedic formulation as claimed in claim 1 having a total of eleven ayurvedic herbal and / or mineral medicines contained in four separate compositions wherein each of the four compositions is selected from a powder composition, a tablet / pill composition, or a capsule composition or syrup composition.
10. The ayurvedic formulation as claimed in claim 1 having a first composition containing Kirat- tikta / Swertia chirata (panchang), Katuka ghan / Picrorrhiza kurroa (root), Jasad bhasm (Oxide of zinc) and Shankh bhasm (Calx of conch shell) and a second composition comprising Lashun /Alium sativum, Jiraka / Cuminum cyminum, Saindhav / Rock salt, Shunthi/Zingiber officinalis, Pippali/ Piper longum,Marich / Piper nigrum and Ramatha /Asafoetida.
11. The ayurvedic formulation as claimed in claim 1 having a first composition containing Kirat- tikta / Swertia chirata (panchang), Katuka ghan / Picrorrhiza kurroa (root), Jasad bhasm (Oxide of zinc) and Shankh bhasm (Calx of conch shell) and a second composition comprising Lashun /Alium sativum, Jiraka / Cuminum cyminum, Saindhav / Rock salt, and a third composition comprising Shunthi/Zingiber officinalis, Pippali/ Piper longum, Marich / Piper nigrum and Ramatha /Asafoetida.

12. The ayurvedic formulation as claimed in claim 1 having a first composition containing Kirat- tikta / Swertia chirata (panchang) and Shankh bhasm (Calx of conch shell), a second composition containing Katuka / Picrorrhiza kurroa (root) and Jasad bhasm (Oxide of zinc) and a third composition comprising Lashun /Alium sativum, Jiraka / Cuminum cyminum, Saindhav / Rock salt, and a fourth composition comprising Shunthi/Zingiber officinalis, Pippali/ Piper longum, Marich / Piper nigrum and Ramatha /Asafoetida.
13. The ayurvedic formulation as claimed in claim 1 wherein content of each herbal and / or mineral medicine is preferably not more than 500 mg, more preferably not more than 400 mg and most preferably not more than 250 mg.
14. The ayurvedic formulation as claimed in claim 1 wherein content of each herbal and / or mineral medicine is from 0.1 % to 25 %, more preferably from 0.5 % to 22.5 % and most preferably, from 1 % to 20 % of the weight of formulation wherein if formulation is capsule, formulation weight is fill weight of capsule excluding weight of capsule shell.
15. The ayurvedic formulation as claimed in claim 2 in the form of a kit.
16. A method of preparing an ayurvedic formulation of claim 1 characterized in having following steps:
i) Dispensing coarse form of herbal medicines based on whether the
formulation has single or 2-3 compositions;
ii) Dispensing separately Bhasma form and fine rock salt.
iii) Grinding each coarse form of medicine to produce finely ground
microfine form;
iv) Subjecting Microfine form of medicines to hydroalcoholic
extraction using cold percolation method to produce hydra-alcoholic
extracts;

v) Subjecting Hydroalcoholic extract of medicines to hydro distillation
in a hydro-distillation machine to obtain dry extract containing little or no
solvent;
vi) If required, drying extracts further to produce dry powder;
vii) Based on individual composition content subjecting to grinding
Dried extracts or dry powder produced from dried extracts, Bhasma form and
fine rock salt, sifting, transferring to a blender and blending to produce
uniform blend or blends of compositions of example 1or 2 or 3 containing
compositions 1-3 and all 11 ingredients in a single composition or 2-3
different compositions.
viii) Optionally blending the blend / blends of step 7 with one or more
inactive ingredients; ix) Filling the blend / blends into a sachet or compressing the blend compressed
into a tablet or filled into a capsule.