Title:  Pharmaceutical formulation useful for the treatment of hepatitis B, hepatitis C and other viral infections of the liver and a process for its preparation

United States Patent:  6,589,570

Issued:  July 8, 2003

Inventors:  Thyagarajan; Sadras Panchatcharam (Chennai, IN)

Assignee:  University of Madras (Chennai, IN)

Appl. No.:  719486

Filed:  December 12, 2000

PCT Filed:  April 10, 2000

PCT NO:  PCT/IN00/00046

371 Date:  December 12, 2000

102(e) Date:  December 12, 2000

PCT PUB.NO.:  WO00/61161

PCT PUB. Date:  October 19, 2000

Abstract

The invention disclosed in this application relates to a pharmaceutical formulation prepared from the biotyped variety of the medicinal plant, Phyllanthus amarus which is useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver with antihepatotoxic and liver cell regenerating potentials and immunomodulating properties. The invention confirms, that when pars of the biotyped variety of the medicinal plant, Phyllanthus amarus are extraceted separately with a polar solvent alone, polar solvent and water in specific ratios and water alone and when such extracts are mixed together, the resultant formulation has all the essential antiviral and biological properties, while the individual polar or aqueous extracts alone does not possess one or more of these properties. The present invention also relates to a process for the preparation of the above said new pharmaceutical formulation with biological and chemical standardisation protocols.

DISCLOSURE OF THE INVENTION

Concept & Hypothesis

It was the feeling that the non-reproducibility of treatment efficacy of the extract of the plant Phyllanthus amarus was due to the absence in the extract of all the under mentioned antiviral and biological properties which are essentially required for the efficient treatment of Hepatitis B (both acute and chronic) Hepatitis C (both acute and chronic) and other related viral infections of the liver.

(i) HBsAg binding property facilitating the inactivation of the virus in circulation ultimately leading to viral clearance

(ii) HBV-DNA polymerase enzyme inhibiting potential, thus acting as anti-viral preventing the multiplication of HBV.

(iii) Reverse Transcriptase enzyme inhibition also required for the initiation of HBV replication.

(iv) inhibition of HBsAg secretion from HBV transinfected liver cells thus possessing activity against virus infected chronic liver disease conditions.

(v) Hepatoprotective and antihepatotoxic properties against the liver cell toxicity brought about by all hepatits viruses (A,B,C,D & E) and other hepatotoxic agents.

(vi) Immunomodulating property to potentiate the immune system of HBV infected patients towards virus clearance and protective antibody (anti HBs) responses.

(vii) HCV replication inhibition as shown by converting HCV-RNA positivity to HCV-RNA negativity thus possessing activity against HCV infected chronic liver disease conditions.

In the light of the above mentioned studies coupled with the above mentioned observations, it was necessary to explain the reasons for non-reproducibility of the clinical efficacy of P. amarus on one hand and to conduct further clinical trials independently in different places using the P. amarus preparation of Thyagarajan. Accordingly clinical trials were conducted on a total of 173 chronic HBV carriers (3 in Chennai (Madras), 1 at Vellore and 1 at Glasgow, UK) subsequent to the earlier to published clinical trials on 98 chronic HBV carriers. The result are presented in Table 3.

In summary, these trials have shown a mean HBsAg clearance rate of 25.6% and mean HBeAg seroconversion rate of 55.3%. It was finally decided to recommend a schedule of 500 mg dosage of P. amarus preparation in capsules given orally for three times daily for six months.

Phyllanthus amarus Treatment in Acute and Chronic Hepatitis C

Available literature in the public domain did not reveal any report on the use of Phyllanthus amarus in the treatment of acute and chronic hepatitis C virus infection which is another major liver pathogen leading to significant morbidity and mortality. Hence the formulation of Phyllanthus amarus envisaged by the present invention was utilised to conduct two clinical trials to treat acute and chronic hepatitis C. Confidential clinical trials have been conducted using the formulation of Phyllanthus amarus in the form of capsules provided by Thyagarajan et al from Chennai.

Since there are no experimental animal/tissue culture models available for testing specific antiviral properties against Hepatitis C virus (HCV), direct studies on human volunteer infected with HCV were conducted. Ethical clearance and informed consents from the study participants were obtained based on the high safety profile of the preparation of P. amarus prepared by the applicants as described above.

Table.4. First Set of Case Studies on the Efficacy of the Preparation of Phyllanthus amarus Treatment on Chronic Hepatitis C Infections Conducted at Glasgow, UK(Eric Walker et al)

1. Patient Having the Date of Birth 07/02/49

Summary: The major finding has been the marked improvement in symptoms when the patient was treated with the preparation of Phyllanthus amarus which symptoms relapsed on 2 occasions when the treatment was withdrawn. The liver enzymes deteriorated when the preparation of Phyllanthus amarus was withdrawn and improved when it was started again. The patient has always been PCR positive. Appearance is of someone who is getting a `liver protective effect` but without elimination of virus. Source was blood transfusion.

29/01/96--Liver biopsy--some inflammation and hepatic fibrosis consistent with chronic hepatitis C infection. Itch, joint pains, lethargy were the main symptoms. Not keen on interferon--Treatment with the preparation of Phyllanthus amarus was started

23/04/96--The weight of the patient increased by 2 kgs. Markedly improved energy and symptoms. Little change in liver enzymes (AST 42,ALT 50). PCR positive

15/05/96 treatment with the preparation of the composition containing Phyllanthus amarus was stopped

17/06/96--Itch, joint pains and lethargy returned (AST 46, ALT 50)

11107/96--Treatment with the composition containing phyllanthus amaraus was restarted

10/09/96--Symptoms were much improved (AST 46, ALT 71)

01/07/97--Still hepatitis C PCR positive(genotype 3). Repeat liver biopsy shows little change (inflammation and fibrosis but no active cirrhotic changes)

05/06/98--weight of the patient was found to be steady and the patient felt well (AST 42, ALT 59)

08/08/98--Treatment with the preparation of Phyllanthus amarus was stopped

31/12198--The patient was found to be depressed and `run down`(AST 62, ALT 75)

211011/99--Treatment with the preparation of Phyllanthus restarted following which a marked improvement in the patient's feeling of well being was observed.

29/12/99--The patient felt well (AST 50, ALT 69), remained PCR positive

2. Patient Having a Date of Birth 20/06/54

Source of HCV infection was intravenous drug use

This patient was one of the first to clear HBsAg and `e` antigen after treatment with the preparation of Phyllanthus tamarus back in 1990--he remains negative (1999) He was not tested for hepatitis C until 1999 (April) when he was found to have antibody but was PCR positive. He remained well.

A second set of studies have been carried out by the inventor. Results confirming the efficacy of the new formulation for the treatment of acute and chronic Hepatitis B & C and other related infections of liver are summarised in Table--6.

THE INVENTION

We observed that if all the above said six essential properties are made available in a single formulation, the resulting formulation will have uniform and stable antiviral and biological potentials which will be beneficial and optimal for the treatment of acute and chronic hepatitis B & C and other viral diseases of the liver. Based on the above findings, the R & D work for the development of a new formulation of P. amarus extract was initiated to find out which formulation will be active significantly against both acute Hepatitis B and against chronic HBV carriers. Accordingly, R & D was directed towards developing such a formulation from the plant Phyllanthus amarus containing all the above said essential characteristics which have not only efficient clinical and biological efficacy against acute and chronic Hepatitis B; acute and chronic Hepatitis C and other viral infections of the liver but also have uniform and stable antiviral and bio active properties.

OBJECTIVES OF THE INVENTION

Accordingly, the main objective of the present invention, therefore, is to provide a pharmaceutical formulation having uniform and stable antiviral and biological efficacy which is useful for the treatment of acute and chronic Hepatitis B. Hepatitis C, chronic HIV carriers and other related viral infections of the liver from the plant, Phyllanthus amarus

Another objective of the present invention is to provide a pharmaceutical formulation useful for the treatment of Hepatitis B, Hepatitis C , chronic HIV carriers and other related viral infections of the liver from the plant Phyllanthus amarus which brings about binding of Hepatitis B surface antigen (HBsAg) of the Hepatitis B virus, thus facilitating the inactivation of the virus in circulation ultimately leading to viral clearance.

Yet another objective of the present invention is to provide a pharmaceutical formulation useful for the treatment of Hepatitis B, Hepatitis C, chronic HIV carriers and other related viral infections of the liver from the plant P. amarus which inhibit the HBV-DNA polymerase enzyme required for the replication for the virus, thus acting as antiviral preventing the multiplication of the virus itself.

Still another objective of the present invention is to provide a pharmaceutical formulation useful for the treatment of Hepatitis B, Hepatitis C, chronic HIV carriers and other related viral infections of the liver from the plant P. amarus which inhibits the Reverse Transcriptase enzyme which is also required for the initiation of HBV replication and is the chief enzyme required for the replication of the AIDS virus, Human Immunodeficiency virus (HIV).

Another objective of the present invention is to provide a process for the preparation of a pharmaceutical formulation useful for the treatment of Hepatitis B, Hepatitis C, chronic HIV carriers and other related viral infections of the liver from the plant P. amarus which is hepatoprotective and also possess antihepatotoxic properties against the liver cell toxicity brought about by all Hepatitis viruses (A,B,C,D & E) and other hepatotoxic agents including chemicals and aflatoxins.

Yet another objective of the present invention is to provide a pharmaceutical formulation useful for the treatment of acute and chronic Hepatitis B; acute and chronic Hepatitis C, chronic HIV carriers and other related viral infections of the liver from the plant P. amarus which is anti-inflammatory and also possesses the property of normalising the transaminase enzymes level indicating antihepatotoxic and liver cell regenerating potentials of the formulation.

Still another objective of the present invention is to provide a pharmaceutical formulation useful for the treatment of acute and chronic Hepatitis B; acute and chronic Hepatitis C , chronic HIV carriers and other related viral infections of the liver from plant P.amarus which inhibited the Hepatitis C virus replication as revealed by conversion from HCV-RNA positivity to HCV-RNA negatitvity in patients treated by the formulation.

Another objective of the present invention is to provide a pharmaceutical formulation useful for the treatment of acute and chronic Hepatitis B; acute and chronic Hepatitis C, chronic HIV carriers and other related viral infections of the liver from plant P.amarus which is immunomodulatory as revealed by increased T-cell proliferation.

Still another objective of the present invention is to provide a pharmaceutical formulation useful for the treatment of acute and chronic Hepatitis B, acute and chronic Hepatitis C and other related viral infections of the liver from the plant P. amarus in which the antiviral and biological activities are uniform.

Yet another objective of the present invention is to provide a process for the preparation of a pharmaceutical formulation useful for the treatment of acute and chronic Hepatitis B acute and chronic Hepatitis C and other related viral infections of the liver from the plant P. amarus with antihepatotoxic, liver cell regenerating and immunomodulating potentials.

The invention is based on our surprising findings that when the parts of the plant Phyllanthus amarus are extracted separately with a polar solvent alone, mixture of polar solvent and water and water alone and when the extracts so obtained are mixed together, the resultant formulation is found to have all the under mentioned properties which are required for the efficient treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver.

(i) HBsAg binding property facilitating the inactivation of the virus in circulation ultimately leading to viral clearance.

(ii) HBV-DNA polymerase enzyme inhibiting potential, thus acting as anti-viral, preventing the multiplication of HBV.

(iii) Reverse Transcriptase enzyme inhibition also required for the initiation of HBV replication.

(iv) Inhibition of HBsAg secretion from HBV transinfected liver cells thus possessing activity against virus infected chronic liver disease conditions.

(v) Hepato-protective, anti-inflammatory, anti-hepatotoxic and liver cell regenerative properties against the liver cell toxicity brought about by all hepatitis viruses (A,BC,D & E) and other hepatotoxic agents.

(vi) Immunomodulating property to potentiate the immune system of HBV infected patients towards virus clearance and protective antibody (anti HBs) responses.

(vii) HCV-replication inhibition as shown by converting HCV-RNA positivity to HCV-RNA negativity thus possessing activity against HCV infected chronic liver disease conditions.

It is observed that the individual extracts of the plant Phyllanthus amarus namely the extract obtained by using a polar solvent alone, extract obtained by using a polar solvent and water and the extract obtained by using water alone, themselves, does not possess all the above said essential characteristics. But the combination of the above mentioned extracts imparts to the resulting formulation all the above mentioned essential properties which are required for the efficient treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver. These properties are acquired by the formulation due to the biological synergism of the different components contained in the individual extracts when combined to form the formulation.

The pharmaceutical formulation of the present invention is not, therefore, a mere admixture of the individual components resulting in the aggregation of the properties of the individual components but is a novel pharmaceutical formulation having biological synergism of all the required efficacious antiviral and biological properties of the components employed.

Accordingly, the present invention provides a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic) Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises

(i) an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone

(ii) an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water and

(iii) an extract of the plant Phyllanthus amarus obtained by using water alone

According to another feature, the present invention provides a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises

(i) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone

(ii) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 20 to 80 and 80 to 20% w/w respectively and

(iii) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using water alone

In another preferred embodiment of the invention, there is provided a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises

(i) 20 to 30% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone

(ii) 20 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 80 to 20% and 20 to 80% w/w respectively

(iii) 20 to 40-% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent and water wherein the solvent to water ratio ranges from 20 to 80% and 80 to 20% w/w respectively and

(iv) 20 to 30% w/w of an extract of the plant Phyllanthus amarus obtained by using water alone

In still another preferred embodiment of the present invention there is provided a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises one part each of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone, an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water and an extract of the plant Phyllanthus amarus using water alone.

In yet another preferred embodiment of the present invention, there is provided a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (bath acute and chronic) and other related viral infections of the liver which comprises one part each of

(i) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone.

(ii) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 20 to 80 and 80 to 20% w/w respectively and

(iii) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using water alone.

In another prefrred embodiment of the present invention, there is provided a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises one part each of

(i) 20 to 30% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone,

(ii) 20 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 80 to 20% and 20 to 80% w/w respectively

(iii) 20 to 40% w/w of an extract of the plant Phyllanthus amarus using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 20 to 80% w/w and 80 to 20% w/w respectively and

(iv) 20 to 30% w/w of an extract obtained using water alone

In still another preferred embodiment of the present invention, there is provided a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises two parts

(i) of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone and one part each of

(ii) of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water and

(iii) an extract of the plant Phyllanthus amarus using water alone

In yet another preferred embodiment of the present invention, there is provided a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other viral infections of the liver which comprises two parts

(i) of 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent and one part each

(ii) of 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 20 to 80% and 80 to 20% w/w respectively

(iii) of 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using water alone

In yet another preferred embodiment of the present invention there is provided a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other viral infections of the liver which comprises two parts

(i) of 20 to 30% W/W of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone and one part each

(ii) of 20 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 80 to 20% and 20 to 80% w/w respectively

(iii) of 20 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of polar solvent and water wherein the ratio of the solvent to water ranges from 20 to 80% w/w and 80 to 20% w/w respectively and

(iv) of 20 to 30% w/w of an extract of the plant Phyllanthus amarus obtained by using water alone.

In yet another preferred embodiment of the invention, there is provided a process for the preparation of pharmaceutical formulation useful for the treatment of Hepatitis B (acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises mixing

(i) an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone

(ii) an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water and

(iii) an extract of the plant Phyllanthus amarus obtained by using water alone

According to another feature the present invention, provides a process for preparing a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises mixing

(i) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone

(ii) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 20 to 80 and 80 to 20% w/w respectively and

(iii) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using water alone

In another preferred embodiment of the invention, there is provided a process for the preparation of a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises mixing

(i) 20 to 30% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone

(ii) 20 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 80 to 20% and 20 to 80% w/w respectively

(iii) 20 to 40-% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent and water wherein the solvent to water ratio ranges from 20 to 80% and 80 to 20% w/w respectively and

(iv) 20 to 30% w/w of an extract of the plant Phyllanthus amarus obtained by using water alone

In still another preferred embodiment of the present invention, there is provided a process for the preparation of pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver comprises mixing one part each of

(i) an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone,

(ii) an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water and

(iii) an extract of the plant Phyllanthus amarus using water alone

In yet another preferred embodiment of the present invention, there is provided a process for the preparation of pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises mixing one part each of

(i) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone.

(ii) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 20 to 80 and 80 to 20% w/w respectively and

(iii) 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using water alone

In another preferred embodiment of the present invention there is provided a process for the preparation of a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises mixing one part each of

(i) 20 to 30% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone,

(ii) 20 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 80 to 20% and 20 to 80% w/w respectively

(iii) 20 to 40% w/w of an extract of the plant Phyllanthus amarus using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 20 to 80% w/w and 80 to 20% w/w respectively and

(iv) 20 to 30% w/w of an extract obtained using water alone

In still another preferred embodiment of the present invention, there is provided a process for the preparation of a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises mixing two parts

(i) of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone and one part each of

(ii) of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water and

(iii) an extract of the plant Phyllanthus amarus using water alone

In yet another preferred embodiment of the present invention, there is provided a process for the preparation of a pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other viral infections of the liver which comprises mixing two parts

(i) of 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a polar solvent and one part each

(ii) of 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 20 to 80% and 80 to 20% w/w respectively

(iii) of 10 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using water alone

In yet another preferred embodiment of the present invention, there is provided a process for the preparation pharmaceutical formulation useful for the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other viral infections of the liver which comprises mixing two parts

(i) of 20 to 30% W/W of an extract of the plant Phyllanthus amarus obtained by using a polar solvent alone and one part each

(ii) of 20 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of a polar solvent and water wherein the ratio of the solvent to water ranges from 80 to 20% and 20 to 80% w/w respectively

(iii) of 20 to 40% w/w of an extract of the plant Phyllanthus amarus obtained by using a mixture of polar solvent and water wherein the ratio of the solvent to water ranges from 20 to 80% w/w and 80 to 20% w/w respectively and

(iv) of 20 to 30% w/w of an extract of the plant Phyllanthus amarus obtained by using water alone.

The present invention also envisages within its scope the use of the pharmaceutical composition of the present invention described above for the preparation of a medicament useful for treating Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver.

The invention further envisages the use of the pharmaceutical composition of the present invention in the treatment of Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver.

Furthermore the present invention also, within its scope includes, the use of the pharmaceutical composition described above for achieving antihepatotoxicity, liver cell regeneration and immunomodulation.

In another embodiment of the present invention there is provided a method of treating Hepatitis B (both acute and chronic), Hepatitis C (both acute and chronic) and other related viral infections of the liver which comprises administering a therapeutically effective dose of the pharmaceutical composition described above.

Extraction of the Components of the Formulation

Parts such as leaves, stems, seeds and roots of the taxonomically identified collections of the plant Phyllanthus amarus were dried and kept in an oven heated to a temperature in the range of 50 to 80 degree C. for a period in the range of 3 to 5 hrs a day. for a period of 3 to 6 successive days. The dried parts of the plants are then powdered. The powder thus obtained is used for the extraction of different components of the preparation of the formulation of the present invention.

Before subjecting the powder for extraction procedures, each batch of powder is subjected for sterility testing to rule out any bacterial or fungal contamination as per standard methods.

One portion of the powder is extracted with polar solvent. The polar solvent employed may be methanol, ethanol, hexane, butanol and the like. The extraction may be carried out at a temperature in the range of 37 to 60 degree C. for a period ranging from 2 hours to 18 hours preferably at a temperature in the range of 37 to 60 degree C. This extract may be used as the component (i) of the pharmaceutical formulation of the present invention.

Another portion of the powder is extracted with a mixture of polar solvent and water. The polar solvent employed may be methanol, ethanol, Hexane and the like. The ratio of the solvent and water used for extraction may range from 20 to 80 and 80 to 20% w/w respectively, The ratio may preferably range from 30 to 50% and 50 to 30% w/w respectively. The extraction may be carried out at a temperature in the range of 4 to 40 degree C. for 2 to 18 hours more preferably at a temperature in the range of 37 to 60 degree C. for 2 to 4 hours. This extract may be used as the component (ii) of the pharmaceutical formulation of the present invention.

Yet another portion of the powder is extracted with water alone. The extraction may be carried out at a temperature in the range of 37 to 60 degree C. for 2 to 18 hours. This extract may be used as the component (iii) of the pharmaceutical formulation of the invention

The extracts so obtained may now be mixed together to obtain the pharmaceutical formulation of the present invention. The mixing may be effected in a vertex mixer or heating mantle and stirring it thoroughly till a homogenous formulation is obtained. The mixing may be effected at a temperature in the range of 37 to 60 degree C. for 15 to 30 minutes.

Claim 1 of 30 Claims

What is claimed is:

1. A pharmaceutical formulation useful for treating acute Hepatitis B, chronic Hepatitis B, acute Hepatitis C, chronic Hepatitis C, and/or other related viral infections of a liver, the pharmaceutical formulation comprising an Extract 1, an Extract 2, and an Extract 3, wherein:

(i) the Extract 1 is an extract of a first amount of a plant Phyllanthus amarus obtained by using a first polar solvent alone, wherein the first polar solvent comprises a polar solvent other than water;

(ii) the Extract 2 is an extract of a second amount of the plant Phyllanthus amarus obtained by using a mixture of a second polar solvent and water alone, wherein the second polar solvent comprises a polar solvent other than water; and

(iii) the Extract 3 is an extract of a third amount of the plant Phyllanthus amarus obtained by using water alone.

____________________________________________
If you want to learn more about this patent, please go directly to the U.S. Patent and Trademark Office Web site to access the full patent.