WO2016025010A1

WO2016025010A1 - A novel broad-spectrum antiviral synergistic pharmaceutical composition for the treatment and prevention of viral infections

Info

Publication number: WO2016025010A1
Application number: PCT/US2014/068451
Authority: WO
Inventors: Martin UGWU
Original assignee: Ugwu Martin
Priority date: 2014-08-14
Filing date: 2014-12-03
Publication date: 2016-02-18
Also published as: WO2016025010A8

Abstract

A broad-spectrum antiviral synergistic pharmaceutical drug composition consisting of a Tetracycline anti-infective agent (doxycycline, minocycline, tetracycline or other derivatives) and Garlic (Allium sativum) extracts (tablets, capsules, mixtures) or aged or fresh bulbs, for the treatment and prevention of viral infections caused by Filoviruses (Ebola Vims & Marburg Vims) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2) is presented. This present invention highlights drug combinations between a tetracycline (doxycycline, minocycline, tetracycline or other derivatives) and Garlic (Allium sativum) extract (tablets, capsules, mixtures) or bulbs, each drug having potent antiviral activities whose combined effect results in synergistic therapeutic effect which is far greater than their additive effects against Filoviruses (Ebola Vims & Marburg Vims) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

Description

A Novel Broad-Spectrum Antiviral Synergistic Pharmaceutical Composition For The Treatment and Prevention Of Viral Infections

DESCRIPTION

FIELD OF INVENTION

Synergistic pharmaceutical drug compositions consisting of a tetracycline (doxycycline, minocycline, tetracycline and other derivatives) and Garlic (Allium sativum) extract (tablets, capsules, bulbs, powder or oil) for the prevention and treatment of infections caused by

Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2) is presented. This present invention relates to drug combinations between a tetracycline (doxycycline, minocycline, tetracycline and other suitable derivatives) and Garlic (Allium sativum) extract (tablets, capsules, mixtures) or bulbs, each drug having potent antiviral activities whose combined effects result in a broad- spectrum synergistic therapeutic effect which is far greater than their additive effects against Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2). More so, it relates to the manufacturing process thereof and the use of this novel drug combination for the prevention, treatment or prophylaxis of several viral infections like those caused Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

It also relates to the use of any of the tetracycline derivatives or salt, solvate, complex, hydrate, isomer, ester, anhydrate, enantiomer, polymorph, pro-drug or any product resulting from chemical modification or synthesis of the afore mentioned and related substances, in

combination with the constituents of garlic (Allicin, Diallyl disulphide, alliin, ajoene and other organosulphur constituents) and or any of their derivatives, or salt, solvate, complex, hydrate, isomer, ester, anhydrate, enantiomer, polymorph , prodrug, or any product resulting from chemical modification or synthesis of the afore mentioned and related substances, in the treatment, prophylaxis or any form of therapeutic intervention against viral infections like those caused Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HTV-2).

BACKGROUND OF THE INVENTION

Ebola Virus

Ebola virus disease (EVD) or Ebola hemorrhagic fever (EHF) is a highly infective disease of humans and other animals caused by the Ebola Virus, a filovirus. It has recorded high mortality rates in several outbreaks since its discovery. The incubation period after contracting the virus is about 2 to 21 days after which symptoms high fever, throbbing headache, sore throat, muscular and body pain are experienced. Usually, gastrointestinal symptoms such as nausea, vomiting, emesis and even diarrhea follow thereafter. Also hepatic and renal functions are impaired. The virus affects platelet function which results in extensive bleeding, both internally and externally as evidenced by coughing out of blood, bloody nasal discharges, petechiae in several areas of the body which indicate bleeding under the skin thus being among the known a viral hemorrhagic infections.

The virus is commonly passed on through contact with infected blood or other bodily fluids of an infected subject and fruit bats have been implicated as carriers of this deadly virus which eventually spread it to humans by contact or when consumed as bush meats. Outbreaks in the human race occurs from transmission of the virus from fruit bats or other animal sources to humans and then may spread between people very fast as a wild raging fire. Usually it is clinically advisable to rule out other diseases since its symptoms are very similar to notable diseases like malaria, cholera and other viral hemorrhagic infections. Diagnosis is usually confirmed by testing for the viral antibodies, viral particles or the virus itself by ELISA method. Male survivors could still be able to transmit the disease through the semen for up to 8 weeks.

Marburg Virus

Marburg virus disease, MVD is also one of the known viral hemorrhagic fevers. It is caused by the Marburg virus, another filovirus, which was first noticed in the 1960s. Just like the EVD, Marburg virus disease causes severe disease in humans and other primates and has recorded high mortality rates during outbreaks.

Since there is no yet known drug treatment for these viral hemorrhagic diseases, quarantine measures, supportive therapy to patients, isolation of suspected cases and preventive measures remain the bases of care in controlling outbreaks.

Presently, there is on ongoing Ebola outbreak in several West African countries and nearby regions and the entire world is under siege with the fear that it could easily to other continents due to inevitable transcontinental human travel. Hence all hands are on deck to find an effective treatment or prophylaxis against this health scourge.

Human Immunodeficiency Virus (HIV)

Retroviruses consist of single-stranded enveloped RNA viruses that replicate in . a host cell through the process of reverse transcription. Some of these include Human Immunodeficiency Virus (HIV), Simian Immunodeficiency Virus (SIV) and Feline Immunodeficiency Virus (FIV).

HIV infection if left untreated or poorly managed may progress to Acquired Immune Deficiency Syndrome (AIDS) which is a medical condition when the immune system is severely compromised to the extent that it can no longer fight infections and thus allows opportunistic infections and certain kinds of neoplasms to evade the subject. HIV infection results by the transmission of the virus in blood or other bodily fluids such as semen, vaginal fluid, or breast milk to an unsuspecting individual.

In the host, HIV progresses to infect and destroy numerous cells of the immune system like macrophages and CD4+ T cells. This retroviral infection potentially leads to low levels of CD4+ T cells through a host of mechanisms, including apoptosis of uninfected bystander cells, direct viral destruction of infected cells, and killing of infected CD4+ T cells by CD8 cytotoxic lymphocytes that recognize infected cells. As the vital CD4+ T cell numbers decline below certain level, cell-mediated immunity is grossly impaired and eventually lost, and hence increasing the vulnerability of the body to become more susceptible to opportunistic infections like candidiasis, cytomegalovirus (CMV) infection, Mycobacterium avium infection,

Pneumocystis pneumonia (PCP), toxoplasmosis and tuberculosis .

Up to 35 million people worldwide are currently living with HIV which has claimed the lives of more than 30 million individuals since it was first reported. Furthermore, in 2011, it recorded 2.5 million newly infected cases globally and its prevalence has transcended across all levels of social strata. Today, HIV epidemic has assumed a health issue in almost all health circles as the treatment, prophylaxis and cure of this retroviral infection still eludes the human race.

The current therapeutic strategy employed to tackle HIV infection is the use of Highly Active Antiretroviral Therapy (HAART). This usually consists of a combination of antiretroviral drugs (ARV) and the modalities involve simultaneous or separate admmistration of combination of drugs, which selectively target different essential points or enzymes of this retroviral life cycle in order to disrupt or halt the viral replication and thus prevent its progression to AIDS.

OBJECT OF THE INVENTION The object of the present invention is to provide a broad-spectrum antiviral synergistic pharmaceutical composition which is a combination therapy to effectively treat or prevent viral infections caused by Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

Another object of the present invention is to provide a therapeutically effective pharmaceutical antiviral composition which would not only be convenient for patient administration but would also promote patient adherence to therapy.

Furthermore, an object of the present invention is to provide a feasible process of manufacturing such a pharmaceutical composition.

Another object of the present invention is to provide a very cost-effective method of prevention, treatment or prophylaxis of viral infections caused by Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

In addition, another object of the present invention is to provide use of the synergistic pharmaceutical composition for the treatment or prophylaxis of viral infections caused by Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

Finally, the object of the present invention is to provide a synergistic pharmaceutical composition which has tolerable side-effect profile for the treatment, prevention and prophylaxis of viral infections caused by Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

SUMMARY OF THE INVENTION

According to one aspect of the present invention there is provided a pharmaceutical composition comprising:

-at least a Tetracycline anti-infective agent, a promoter of the expression of zinc-finger antiviral protein (ZAP) which inhibits viruses by significantly enhancing the degradation of specific viral mRNAs and

-at least Garlic, a broad spectrum antimicrobial agent and an activator of ZAP for maximal activity through the promotion of glucose synthase kinase -3 beta (GSK-3 beta) phosphorylation activities as ZAP requires phosphorylation by GSK-3 beta for maximal antiviral activity. And coupled with other antiviral activities, works synergistically with a tetracycline anti-infective in the elimination of Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-l and HIV-2). Overexpression of ZAP in infected cells renders them resistant to some viruses in a ZAP expression level-dependent manner, whereas depletion of endogenous ZAP enhanced progression of the viral infection.

It has been elucidated that the activation of GSK-3 β is achieved through S-Nitrosylation with nitric oxide which is indirectly generated in vivo by garlic through activation of nitric oxide synthase (Shimizu et al, 2007).This increased phosphorylation of ZAP due to increased activity of GSK-3 beta results in a far much more potent antiviral activity of this novel antiviral synergistic drug combination against Filoviruses (Ebola Virus & Marburg Virus) and

Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

Consequently, the phosphorylation of ZAP by GSK-3 beta, coupled with other complementary antiviral mechanisms, significantly potentiates its antiviral activity to the extent of exerting a highly significant synergistic activity that could be employed in the prevention, treatment or prophylaxis of diseases caused by Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

According to a further aspect of the present invention, there is provided the pharmaceutical composition for use in the prevention, treatment or prophylaxis of viral infections caused by Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

According to another aspect of the present invention, there is provided the use of the

pharmaceutical composition in the manufacture of a medicament for the prevention, treatment or prophylaxis of viral infections caused Filoviruses (Ebola Virus & Marburg Virus) and

Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

According to another aspect of the present invention, there is provided a method of preventing or treating viral infections caused by Filoviruses (Ebola Virus & Marburg Virus) and

Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2) wherein the method comprises administering the pharmaceutical composition of the present invention to a subject in need thereof.

According to another aspect of the present invention, there is provided the use of this pharmaceutical composition as potent antioxidant therapeutic modality in the treatment and prevention of various medical conditions related to infectious diseases, neurology and oncology.

DETAILED DESCRIPTION

As we all know, there is an urgent need to discover, develop and formulate an effective and tolerable drug or pharmaceutical composition for the treatment of viral infections caused by Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HrV-1 and HIV-2) which would not only be convenient for patient administration but would also support patient adherence to the therapy.

The inventor of the present invention has now amazingly found combinations of a tetracycline (doxycycline, minocycline, tetracycline and other derivatives) and Garlic (Allium sativum) extract (tablets, capsules, bulbs, powder or oil) for the treatment, prevention and prophylaxis of viral infections caused by Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

Though tetracyclines are well known for their antibacterial activity via the inhibition of protein synthesis, specifically by binding to the 30S nbosomal subunit in the messenger RNA (mRNA), they are indeed very dynamic anti-infective molecules which have been proven to be very efficacious in diverse fields of research such as virology, neurology, and oncology. Tetracyclines exert antiviral effects against filoviruses and retroviruses (Fuoco, 1999).

Garlic is a naturally occurring armamentarium in the fight against infections caused by various micro-organisms like bacteria, viruses and fungi. It is widely known in traditional medicine for its usefulness in the treatment of various infections. Garlic (Allium sativum) has been shown to have antiviral activities and the extract or bulbs usually consists of diallyl thiosulfinate (allicin), allyl methyl thiosulfinate, methyl allyl thiosulfinate, ajoene, alliin, deoxyalliin, diallyl disulfide, diallyl trisulfide and other organosulphur compounds.

The administration of a tetracycline antibiotic promotes the expression of zinc-finger antiviral protein (ZAP) which has been described as a host protein conferring resistance to retroviral infection. Expression of (ZAP) in cells has been found to inhibit the replication of viruses such as like retroviruses, filoviruses and herpes viruses.

The present invention provides a pharmaceutical composition comprising at least one tetracycline, a promoter of the expression of zinc-finger antiviral protein (ZAP) which inhibits the replication of retroviruses by significantly enhancing the degradation of specific viral mRNAs and at least garlic (Allium sativum) extract, a naturally occurring plant with known antiviral properties which may also promote and induce the activity of Glucose synthase kinase - 3 beta (GSK-3 beta), a biomolecule with several metabolic and immunologic functions.

Overexpression of ZAP in infected cells renders them resistant to retroviral infections in a ZAP expression level-dependent manner, whereas depletion of endogenous ZAP enhances the progression of the viral infection. Phosphorylation of ZAP by GSK3 enhances the antiviral activity of ZAP against filoviruses and retroviruses (Fuoco, 2012).

Several researches on the interaction between ZAP and GSK-3 beta strongly suggest that phosphorylation is required for optimal activity of ZAP. This has been supported by the fact that the overexpression of GSK3 in ZAP cells, and thus the phosphorylation of ZAP specifically at the S5 to SI sites, significantly increased ZAP activity against viruses such as retroviruses, filoviruses and herpes viruses (Sun et al, 2012). It has been elucidated that the activation of GSK-3 is achieved through S-Nitrosylation with nitric oxide (Shimizu et al, 2007).

This increased phosphorylation of ZAP due to increased activity of GSK-3 beta results in a far much more potent antiviral activity of this novel antiviral synergistic drug combination against sensitive viruses.

Consequently, the phosphorylation of ZAP by GSK-3 beta significantly potentiates its antiviral activity to the extent of exerting a pronounced synergistic interaction that could be employed in the prevention, treatment or prophylaxis of diseases caused by Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HTV-2).

The viral RNA polymerase is another site of action of this novel synergistic drug combination in viral replication process. It has been discovered that the main antimicrobial effect of Garlic, which is rich is several organosulphur substances like Ajoene and allicin, is based on its chemical reaction with thiol groups of various enzymes like the viral RNA polymerase, which plays a vital role in the replication and transcription process (Ankri and Mirelman, 1999). This reaction incapacitates this enzyme and disrupts its functions hence inhibits the replication process and ultimately has a negative effect of their virulence.

Furthermore, the mRNAs is a vital site of action by this novel synergistic drug combination The key mechanism of action of doxycycline as an antiviral agent stems from the fact that its administration promotes the expression of Zinc-finger Antiviral Protein (ZAP) which has been described as a host protein conferring resistance to Ebola, HIV-1 and Sindbis viral infections (Guo et al, 2012).

In addition, it was been unveiled that Doxycycline, which induced the expression of Zinc-finger Antiviral Protein (ZAP), decreased the activity of an EBOV replicon system by up to 95%. It was concluded that ZAP exhibits antiviral activity against filoviruses, by decreasing the level of viral mRNA (Muller et al, 2007).

Expression of (ZAP) in cells has been found to inhibit the replication of viruses such as filoviruses, retroviruses and herpes viruses by significantly enhancing the degradation of specific viral mRNAs by recruiting cellular mRNA degradation machinery (Zhu et al, 2011). This has been supported by investigative results in both human and rat ZAP against the propagation of these viruses.

Specifically, ZAP targets the multiply spliced mRNAs for degradation, selectively recruits cellular poly(A)-specific ribonuclease (PARN) to shorten the poly(A) tail of target viral mRNA and recruits the RNA exosome to degrade the RNA body from the 3' end . In addition, ZAP recruits cellular decapping complex through its cofactor RNA helicase p72 to initiate degradation of the target viral mRNA from the 5' end. (Zhu et al, 2011). In effect, this novel antiviral synergistic drug combination provides a sequential blockade in viral replication process which supports its synergistic antiviral activities for the treatment of viral infections caused by filoviruses and retroviruses.

Another interesting aspect of this novel synergistic drug combination lies in the fact that it has several other antiviral mechanisms that would aid in the elimination of filoviruses and retroviruses.

One of its constituents, garlic, promotes and enhances the activities of GSK-3 beta which then phosphorylates ZAP whose expression in cells was induced by the administration of a tetracycline. Phosphorylation of ZAP by GSK-3 β enhances the antiviral activity of ZAP against viruses such as retroviruses, filoviruses and herpes viruses. In fact, several research findings on the interaction between ZAP and GSK-3 beta strongly suggest that phosphorylation is required for optimal activity of ZAP. This has been supported by the fact that the overexpression of GSK-3 β in ZAP cells, and thus the phosphorylation of ZAP specifically at the S5 to SI sites, significantly increased ZAP activity against viruses like filoviruses (Sun et al, 2012). It has been elucidated that the activation of GSK-3 β is achieved in vivo through S-Nitrosylation reaction with nitric oxide (Shimizu et al, 2007).

This increased phosphorylation of ZAP due to increased activity of GSK-3 beta results in a far much more potent antiviral activity of this novel antiviral synergistic drug combination against Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

This novel synergistic drug combination has the ability to stimulate the generation of nitric oxide, in vivo through the conversion of garlic's constituent, arginine to citrulline, which also accounts for its several antiviral properties (Singla et al, 2011). In addition, fresh and aged garlic extracts have been known to enhance the production of nitric oxide by promoting the activity of nitric oxide synthase (Das et al, 1995).

Nitric oxide has been well studied and accepted by virologists as one of the potent defenses for inhibiting viral replication through various mechanisms which include:

- The Nitric Oxide Pathway Provides Innate Antiviral Protection in Conjunction with the Type I Interferon Pathway in Fibroblasts (Mehta et al, 2012).

- Nitric oxide inhibits HIV-1 replication in human astrocytoma cells (Persichini, 1999)

- Inhibition of viral proteases by S-Nitrosylation (Saura et al, 1999)

- Indirect potentiation of ZAP for maximal activity through the activation of GSK3b by S- Nitrosylation (Shimizu et al, 2007) - Inhibition of cathepsin B and cathepsin L proteases which are required for the cleavage of the ebola GP (Schornberg et al, 2006)

Furthermore, this novel synergistic drug combination which has a garlic constituent, has several identifiable organosulphur constituents which are able to interact with thiol groups in viral R A polymerase and in the helical structure of the viral genome thereby distorting the viral replication and transcription process (Singla et al, 2011).

In addition, this novel drug composition also presents a unique potent antioxidant modality for therapeutic interventions in antimicrobial pharmacotherapy and disease states in neurology and oncology which may present with oxidative stress of different degrees ( ladna et al, 2012).

It will be grateful that the respective therapeutic agents may be administered simultaneously or separately either in the same or different pharmaceutical compositions. If there is separate administration, it will also be appreciated that the subsequently administered therapeutic agents should be administered to a patient within a time scale so as to achieve, or optimize, synergistic therapeutic effect of such a combined pharmaceutical preparation.

The term "pharmaceutical composition" includes tablets, capsules (filled with powders, pellets, beads, mini-tablets, pills, micro-pellets, small tablet units, Multiple Units Pellets System

(MUPS), disintegrating tablets, dispersible tablets, granules, and microspheres, multiparticulates), sachets (filled with powders, pellets, beads, mini-tablets, pills, micro-pellets, small tablet units, MUPS, disintegrating tablets, dispersible tablets, granules, and microspheres, multiparticulates) and sprinkles, however, other dosage forms such as controlled release formulations, lyophilized formulations, modified release formulations, delayed release formulations, extended release formulations, pulsatile release formulations, dual release formulations and the like; liquid dosage form (liquids, suspensions, solutions, emulsions, micro-emulsions, sprays, spot-on), injection preparations, nano formulation etc. may also be included under the control of the invention. Suitable excipients may be used for formulating the various dosage forms according to the present invention.

The term tetracycline used throughout the description and claims are used in a broad sense to include not only the active ingredient per se but also pharmaceutically acceptable derivatives thereof. Suitable derivatives include pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, pharmaceutically acceptable isomers, pharmaceutically acceptable esters, pharmaceutically acceptable anhydrates, pharmaceutically acceptable enantiomers, pharmaceutically acceptable polymorphs, pharmaceutically acceptable prodrugs, pharmaceutically acceptable tautomers and/or pharmaceutically acceptable complexes thereof and combinations thereof.

Examples of suitable tetracyclines include but are not limited to doxycycline, minocycline, and tetracycline. The preferred tetracycline anti-infective agent in this drug composition is Doxycycline. It has excellent oral bioavailability following administration, though the IV dosage form should be used in reclined patients. It is mostly eliminated non-renally hence could be of great use in renal- impaired patients. More so, its major route of excretion is via the intestines hence would further help in the complete clearance of any latent viruses in the large intestine and rectum.

Doxycycline, which is more conveniently dosed twice daily for better patient compliance, enjoys excellent distribution throughout the body tissues and fluids and also achieves adequate therapeutic concentrations in even sanctuary sites hence would ensure optimal elimination of viruses from the body when combined with garlic.

The dose of doxycycline is in the range of 100 to 200 mg, or to be determined, and adjusted accordingly.

The dose of minocycline is in the range of 50 to 200 mg. or to be determined, and adjusted accordingly.

The dose of tetracycline is in the range 125 to 2000mg. or to be determined, and adjusted accordingly

The Garlic (Allium sativum) extract used may be in the form of, but not limited to

pharmaceutical dosage forms like tablets, capsules, granules, powders, elixirs, liquids, or fresh bulbs or aged-garlic forms. Garlic extract and the fresh bulbs have wide dose range, or to be determined, and adjusted accordingly.

Preferably, the pharmaceutical composition comprises doxycycline and garlic extract or fresh bulbs optionally with one or more pharmaceutically suitable and acceptable excipients.

Alternatively, the pharmaceutical composition comprises minocycline and garlic extract or fresh bulbs optionally with one or more pharmaceutically suitable and acceptable excipients.

Alternatively, the pharmaceutical composition comprises tetracycline and garlic extract or fresh bulbs optionally with one or more pharmaceutically suitable and acceptable excipients.

Suitably, the pharmaceutical composition according to the present invention could be in solid dosage, semi-solid and liquid dosage forms, and other dosage forms fall under the ambit of the invention.

Consequently, the synergistic pharmaceutical composition according to the present invention may also be presented in the form of a kit comprising at least one between a tetracycline (doxycycline, minocycline, tetracycline or other derivatives) and Garlic (Allium sativum) extract or bulbs and provides the patient with his daily regimen of drugs in a single package. This packaging facilitates the patient in receiving the drug regimen of the entire day in a single package which also enables the patient to avoid carrying of numerous medications and also confirm if the same are administered. The kit composition has a merit over the other packaged dosage forms in that the patient always has access to the set of instructions for administration contained in the kit. The inclusion of a set of instructions for appropriate medication

administration has been shown to improve patient compliance.

Preferably, the present invention provides a pharmaceutical antiviral composition consisting doxycycline and garlic extract or bulbs in a kit form.

Preferably, the present invention provides a pharmaceutical antiviral composition consisting minocycline and garlic extract or bulbs in a kit form.

Preferably, the present invention provides a pharmaceutical antiviral composition consisting tetracycline and garlic extract or bulbs in a kit form.

Alternatively, the synergistic pharmaceutical composition in a kit form may comprise a separate unit dosage form of doxycycline, a separate unit dosage form of garlic extract or bulbs, separate unit dosage form of minocycline and separate unit dosage form of garlic extract or bulbs.

The dosage forms such as mini-tablets or granules filled in hard gelatin capsules or sachets may be directly administered or can be administered by sprinkling the mini-tablet or granules on regular meals. Alternatively, the mim-tablets or granules filled in hard gelatin capsules or sachets may be administered with liquid or semi-solid beverages such as but not limited to, juices, water.

The dosage forms such as mini-tablets or granules according to the present invention may also optionally be suitably coated. Preferably, mim-tablets or granules according to the present invention may be film-coated, enteric coated or have other extended release formulations.

Furthermore, these dosage forms may be seal-coated, film-coated and further filled in hard gelatin capsules or sachets.

The synergistic pharmaceutical composition may be administered as a single layered or bilayered or multilayered tablet wherein each layer may or may not contain drug/drugs along with pharmaceutically acceptable excipients which are then compressed to provide either a single layered, bilayered or multilayered tablet. Suitable and acceptable excipients may be used for formulating the various dosage forms.

The term excipients used herein includes one or more of pharmaceutically acceptable and suitable ingredients but are not limited to carriers, diluents or fillers, colorants, binders, lubricants, glidants, flavoring agents and disintegrants. PATENT CITATIONS

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Claims

Claims:

1. A broad-spectrum antiviral synergistic pharmaceutical composition comprising at least a tetracycline (doxycycline, minocycline, tetracycline or other derivatives) an anti-infective agent and Garlic (Allium sativum) extracts (tablets, capsules, mixtures, powders or oils) or bulbs, and optionally one or more pharmaceutically acceptable and suitable excipients for the treatment or prophylaxis of viral infections caused by Filoviruses (Ebola Virus & Marburg Virus) and Retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2)

2. The pharmaceutical composition according to claim 1, comprising at least a tetracycline (doxycycline, minocycline, tetracycline or other derivatives), an anti-infective agent and Garlic (Allium sativum) extracts (tablets, capsules, mixtures, powders or oils) or aged or fresh bulbs, in the form of a pharmaceutically acceptable and suitable derivative thereof.

3. The pharmaceutical composition according to claim 2, wherein the pharmaceutically acceptable and suitable derivative is a salt, solvate, complex, hydrate, isomer, ester, tautomer, anhydrate, enantiomer, polymorph, prodrug or any product resulting from chemical modification or synthesis of the afore mentioned and related substances.

4. The pharmaceutical composition according to any preceding claim, wherein one comprising at least a tetracycline (doxycycline, minocycline, tetracycline or other derivatives), an anti-infective agent.

5. The synergistic pharmaceutical composition according to any preceding claim, wherein comprising at least Garlic (Allium sativum) extracts (tablets, capsules, mixtures, powders or oils) or bulbs.

6. The pharmaceutical composition according to any preceding claim, comprising a tetracycline (doxycycline, minocycline, tetracycline or other derivatives) an anti-infective agent and Garlic (Allium sativum) extracts (tablets, capsules, mixtures, powders or oils) or bulbs.

7. The pharmaceutical composition according to any one of claims 1 to 5, comprising at least a tetracycline (doxycycline, minocycline, tetracycline or other derivatives) an anti-infective agent and Garlic (Allium sativum) extracts (tablets, capsules, mixtures, powders or oils) or bulbs.

8. The synergistic pharmaceutical composition according to any preceding claim, wherein the composition further comprises one or more pharmaceutically acceptable and suitable excipients selected from carriers, diluents, fillers, binders, lubricants, colorants, glidants, disintegrants, bulking agents, flavoring agents or any combination thereof.

9. The pharmaceutical composition according to any preceding claim, wherein the composition is in the form of a tablet, mini-tablet, granules, sprinkles, capsules, sachets, powders, pellets, a liquid dosage form, injection preparation or in the form of a kit.

10. The pharmaceutical composition according to any preceding claim, wherein the composition is a hot melt extruded pharmaceutical formulation.

11. The pharmaceutical composition according to claim 10, wherein the composition further comprises at least one water soluble polymer and/or water swellable polymer and/or water insoluble polymer, plasticizer or any combination thereof.

12. A process for preparing a pharmaceutical composition as claimed in any one of claims 1 to 11, wherein the process comprises admixing the at least comprising at least a tetracycline (doxycycline, minocycline, tetracycline or other derivatives) an anti-infective agent and Garlic (Allium sativum) extracts (tablets, capsules, mixtures, powders or oils) or bulbs, and optionally pharmaceutically acceptable and suitable excipients.

13. The process according to claim 12, comprising the steps of: (a) sifting and dry mixing the at least comprising a tetracycline (doxycycline, minocycline, tetracycline or other derivatives) an anti-infective agent and Garlic (Allium sativum) extracts (tablets, capsules, mixtures, powders or oils) or bulbs, and optionally pharmaceutically acceptable and suitable excipients, (b) forming granules of the at least a tetracycline (doxycycline, minocycline, tetracycline or other

derivatives) an anti-infective agent and Garlic (Allium sativum) extracts (tablets, capsules, mixtures, powders or oils) or bulbs, optionally pharmaceutically acceptable and suitable excipients.

14. The process according to claim 13, wherein the granules are either compressed to form a single layered tablet, bi/multi layered tablet or a mini-tablet; or filled into hard gelatin capsules or sachets.

15. The pharmaceutical composition according to any one of claims 1 to 11, for use in the prevention, treatment or prophylaxis viral infections like those caused by filoviruses (ebola virus & marburg virus) and retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2).

16. Use of a pharmaceutical composition according to any one of claims 1 to 11, in the manufacture of a medicament for the prevention, treatment or prophylaxis of diseases caused by filoviruses (ebola Virus & marburg virus) and retroviruses (Human Immunodeficiency Viruses (HIV-1 and HIV-2) and other diseases caused by sensitive microbes. antioxidant, in the manufacture of a medicament for the prevention, treatment or therapeutic modality for various disease conditions in neurology and oncology aspects of medicine.

18. A method of preventing or treating diseases caused by Filovimses (Ebola Vims & Marburg Vims) and Retro vimses (Human Immunodeficiency Vimses (HIV-1 and HIV-2) wherein the method comprises administering a pharmaceutical composition according to any one of the claims 1 to 11 to a subject in need thereof.

19. The method according to claim 18, wherein the preventing or treating viral diseases caused by Filovimses (Ebola Vims & Marburg Vims) and Retrovimses (Human Immunodeficiency Vimses (HIV- 1 and HIV-2) such as is the preventing, treating of, or any sort of therapeutic intervening against infections caused by these vimses.