WO2022079496A1

WO2022079496A1 - A pharmaceutical composition of nitazoxanide, albendazole and pyrantel and method thereof

Info

Publication number: WO2022079496A1
Application number: PCT/IB2021/050773
Authority: WO
Inventors: Chander Mohan Negi
Original assignee: Menschlich Healthcare Opc Private Limited
Priority date: 2020-10-13
Filing date: 2021-02-01
Publication date: 2022-04-21

Abstract

A pharmaceutical composition for treating Covid-19 virus comprising a therapeutically effective amount of a nitazoxanide or its pharmaceutically acceptable salts thereof, an albendazole or its pharmaceutically acceptable salts thereof and a pyrantel or its pharmaceutically acceptable salts thereof is disclosed. The pharmaceutical composition comprises the nitazoxanide in the ratio of 0.05% to 50% w/v, the albendazole 0.02% to 30% w/v and the pyrantel in the ratio of 0.0125% to 20% w/v. The composition is found to be effective for the treatment of COVID -19 (SARS-CoV2). The pharmaceutical composition of nitazoxanide, albendazole and pyrantel has been found to be effective and is unexpectedly well tolerated with a low rate of side-effects, and equally high cure-rates than in comparable treatments.

Description

A PHARMACEUTICAL COMPOSITION OF NITAZOXANIDE, ALBENDAZOLE AND PYRANTEL AND METHOD THEREOF EARLIEST PRIORITY DATE: This Application claims priority from a Provisional patent application filed in India having Patent Application No. 202021044508, filed on October 13, 2020, and titled “A PHARMACEUTICAL COMPOSITION”. FIELD OF INVENTION Embodiments of a present disclosure relate to a technical field of pharmaceutical composition. More particularly, the present disclosure relates to pharmaceutical composition that includes a synergistic combination of Nitaoxanide, Albendazole and Pyrantel. BACKGROUND It is known that coronavirus disease (COVID-19) is an infectious disease caused by a newly discovered coronavirus. The disease emerged first in December 2019 in Wuhan, Hubei, China. This deadly disease rapidly spread to almost 188 countries around the world, with a mortality rate of approximately 5%. The outbreak was declared a Public health Emergency of International Concern by the World Health Organization (WHO) in January 2020 and was recognized as a pandemic in March 2020. As of 6 October 2020, more than 35.4 million cases have been reported in 188 Countries and territories worldwide, although the true number of cases is likely to be much higher. As per the record more than 1.04 million deaths attributed to COVID-19 as on date. A novel coronavirus, (SARS-CoV), was identified as the etiological agent of SARS. SARS-CoV is an enveloped, positive-strand RNA virus. Its large RNA genome is approximately 30000 nucleotides in length and encodes a nonstructural replicase complex and structural proteins, including spike (S), envelope (E), membrane (M) and nucleocapsid (N) proteins. Spike protein is the envelope protein responsible for invasion of host cells. It has been found that COVID-19 positive individual may remain asymptomatic or show symptoms that may be common, mild in nature or severe. Most common symptoms of COVID-19 positive range from fever, dry cough, tiredness, etc., ranging to less common symptoms such as body aches and pains, sore throat, diarrhea, conjunctivitis, severe headache and loss of taste or smell and appearance of rash on skin, or discoloration of fingers or toes. The disease COVID-19 can spread from person to person through small droplets from the nose or mouth land on objects and surfaces around the person which are spread when a person with COVID-19 coughs or exhales. The disease COVID-19 present as atypical pneumonia that often leads to acute respiratory distress syndrome and respiratory failure, the main cause of death. This unusual infectious disease has become a major threat to public health and social stability since its outbreak. COVID-19 is fatal if the affected individual develops serious symptoms such as difficulty in breathing or shortness of breath, chest pain or pressure and loss of speech or movement. One of the major factors contributing to the continued presence of Covid-19 is that there is no promising therapy available for the disease. In case COVID-19 is detected early on and the affected person takes the due care it has been seen that mild and less serious symptoms can be controlled and overcome. As most of COVID-19 early symptoms are curable, an effective medicine can be formulated with use of existing pharmaceutical salts. The traditional drug development process is slow and not very effective against emerging public health threats such as current one. The manufacturing of the new drug possesses various challenges. Whenever the drug starts new clinical trials, there are lots of problems that needs to be overcome, such as drug safety, patient selection, trial dose and other issues. Even after the drug has been approved by the regulatory authorities and sales on the market, they still may possibly face the situation of the poor drug response in patients. The current invention is comprised of such novel composition which has been effectively selected and is proven to be effective in treatment of COVID 19(SARS-CoV2). Hence, there is a need for a pharmaceutical composition and process for preparation of the same to satisfy the existing needs, as well as others, and generally overcomes the deficiencies found in the prior art. OBJECTIVE OF THE INVENTION In accordance with an embodiment of the present invention, it is the object of the present invention to provide an effective pharmaceutical composition for treatment of COVID-19 (SARS-CoV-2). In accordance with an embodiment of the present invention, it is another object of the present invention to provide a method of production of a pharmaceutical composition comprising Nitazoxanide, Albendazole and Pyrantel. In accordance with an embodiment of the present invention, it is further object of the present invention to provide a composition comprising a simple, easy to manufacture and therapeutically effective dosage regime. SUMMARY OF THE INVENTION In accordance with an embodiment of the present invention, a pharmaceutical composition for treating Covid-19 virus comprising a therapeutically effective amount of a nitazoxanide or its pharmaceutically acceptable salts thereof, an albendazole or its pharmaceutically acceptable salts thereof and a pyrantel or its pharmaceutically acceptable salts thereof. In accordance with an embodiment of the present invention, the pharmaceutical composition comprises the nitazoxanide in the ratio of 0.05% to 50% w/v. In accordance with an embodiment of the present invention, the pharmaceutical composition comprises the albendazole in the ratio of 0.02% to 30% w/v. In accordance with an embodiment of the present invention, the pharmaceutical composition comprises pyrantel in the ratio of 0.0125% to 20% w/v. In accordance with an embodiment of the present invention, the pharmaceutical composition comprises at least one of pharmaceutically acceptable excipient. In accordance with an embodiment of the present invention, the pharmaceutical composition comprises at least one of pharmaceutically acceptable carrier. In accordance with an embodiment of the present invention, the pharmaceutical composition is a granule, tablet, capsule, liquid, or mixture pellets. In accordance with an embodiment of the present invention, the pharmaceutical composition can be administered as oral, parental, topical or any In accordance with an embodiment of the present invention, the combined preparation comprises of one or more dosage units of a nitazoxanide or its pharmaceutically acceptable salts thereof, an albendazole or its pharmaceutically acceptable salts thereof and a pyrantel or its pharmaceutically acceptable salts thereof. To further clarify the advantages and features of the present invention, a more particular description of the invention will follow by reference to specific embodiments thereof, which are illustrated in the appended figures. It is to be appreciated that these figures depict only typical embodiments of the invention and are therefore not to be considered limiting in scope. The invention will be described and explained with additional specificity and detail with the appended figures. BRIEF DESCRIPTION OF THE DRAWINGS The disclosure will be described and explained with additional specificity and detail with the accompanying figures in which: FIG. 1 is a graph illustrating an anti-viral testing results of the pharmaceutical composition comprising nitazoxanide, albendazole and pyrantel in accordance with an embodiment of the present disclosure. Further, those skilled in the art will appreciate that elements in the figures are illustrated for simplicity and may not have necessarily been drawn to scale. Furthermore, in terms of the construction of the device, one or more components of the device may have been represented in the figures by conventional symbols, and the figures may show only those specific details that are pertinent to understanding the embodiments of the present disclosure so as not to obscure the figures with details that will be readily apparent to those skilled in the art having the benefit of the description herein. DETAILED DESCRIPTION For the purpose of promoting an understanding of the principles of the disclosure, reference will now be made to the embodiment illustrated in the figures and specific language will be used to describe them. It will nevertheless be understood that no limitation of the scope of the disclosure is thereby intended. Such alterations and further modifications in the illustrated system, and such further applications of the principles of the disclosure as would normally occur to those skilled in the art are to be construed as being within the scope of the present disclosure. The terms "comprises", "comprising", or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a process or method that comprises a list of steps does not include only those steps but may include other steps not expressly listed or inherent to such a process or method. Similarly, one or more components, compounds, and ingredients preceded by "comprises... a" does not, without more constraints, preclude the existence of other components or compounds or ingredients or additional components. Appearances of the phrase "in an embodiment", "in another embodiment" and similar language throughout this specification may, but not necessarily do, all refer to the same embodiment. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. The system, methods, and examples provided herein are only illustrative and not intended to be limiting. In the following specification and the claims, reference will be made to a number of terms, which shall be defined to have the following meanings. The singular forms “a”, “an”, and “the” include plural references unless the context clearly dictates otherwise. The COVID-19(SARS-CoV-2) is an ongoing pandemic of Corona virus disease 2019(COVID-19). It is an infectious disease caused by newly discovered Severe acute respiratory syndrome Corona virus 2 (SARS-CoV-2). The disease (SARS- CoV-2) can spread from person to person through small droplets from the nose or mouth land on objects and surfaces around the person which are spread when a person with COVID-19 coughs or exhales. The SARS presents as atypical pneumonia that often leads to acute respiratory distress syndrome and respiratory failure, the main cause of death. This unusual infectious disease has become a major threat to public health and social stability since its outbreak. Despite massive international attempts to stop this COVID & SARS-CoV2, it remains a serious pandemic disease around the world. One of the major factors contributing to the continued presence of SARS-CoV2 is that there is no promising therapy available for the disease and the traditional drug development processes are slow and not very effective against emerging public health threats such as the current SARS-CoV-2 coronavirus outbreak. Embodiments of the present invention relates to a pharmaceutical composition for treating Covid-19 virus comprising a therapeutically effective amount of a nitazoxanide or its pharmaceutically acceptable salts thereof, an albendazole or its pharmaceutically acceptable salts thereof and a pyrantel or its pharmaceutically acceptable salts thereof. In accordance with an embodiment of the present invention, the pharmaceutical composition comprises of the nitazoxanide in the ratio of 0.05% to 50% w/v. In accordance with an embodiment of the present invention, the pharmaceutical composition comprises of the albendazole in the ratio of 0.02% to 30% w/v. In accordance with an embodiment of the present invention, the pharmaceutical composition comprises of the pyrantel in the ratio of 0.0125% to 20% w/v. Chemically, nitazoxanide is the prototype member of thiazolides, a class of drugs which are synthetic nitrothiazolyl-salicylamide derivates with broad-spectrum anti-parasitic and broad-spectrum antiviral activity suggests that it possesses efficacy in treating a number of viral infections as well. When these drugs nitazoxanide, albendazole and pyrantel are used alone not in combination, despite, their excellent intrinsic antimalarial activity, they have not shown the any effective results for the treatment against SARS-CoV2 infection. According to an embodiment of the present invention, the nitazoxanide combines synergistically with other classes of anthelmintic drugs i.e., albendazole and pyrantel. This combination of nitazoxanide, albendazole and pyrantel exhibited promising In-Vitro antiviral activity against SARS-CoV2. Therefore, this promising In-Vitro results renders the nitazoxanide, albendazole and pyrantel combination a good candidate for its use in drug combination therapy for SARS- CoV2 infections. EXPERIMENTAL RESULTS FIG. 1 is a graph illustrating an anti-viral testing results of the pharmaceutical composition comprising nitazoxanide, albendazole and pyrantel in accordance with an embodiment of the present disclosure. According to an embodiment of the present invention, the assay details of anti- viral test include:

According to an embodiment of the present invention, the drug combination showed anti-viral activity from 6.25 μM and 100 & 200 μM concentration of the % of viral reduction is ~93%. The viral particles reduced from 10^6.1 to 10^4.9. The experiment was done in duplicates and the values were averaged to calculate % viral reduction. According to an embodiment of the present invention, the regression equation for viral particles Vs Ct value of N-gene specific to SARS-CoV2 virus (y = -4.9474x + 39.723, R² = 0.9964) (X= Number of viral particles, y=Ct value). The number of viral particles are calculated using X= (39.723-Ct N-gene @ different time points.) /4.9474.

According to an embodiment of the present invention, the broad scheme of assay includes as represented below, wherein % survival is determined by cytopathic effects or by viral enumeration in the cell supernatant by RT-PCR:

According to an embodiment of the present invention, the materials for the assay includes but are not limited to: SARS COVID-2 viral isolate (patient origin), Host cell line: Vero (African green monkey kidney cells, Vero (ATCC® CCL-81™) Media and reagent: a. Cell Culture Media (Eagle’s Minimum Essential Media b.10% Fetal Bovine Serum (FBS) c. Phosphate Buffered Saline (PBS) According to an embodiment of the present invention, the assay protocol includes labelling all petri dishes, dilution tube racks, and host-containing apparatus with the following information: virus, host, test agent, and project number. According to an embodiment of the present invention, briefly, a flask of Vero cell grown in cell culture media containing 10% fetal bovine serum (FBS) will be used. Then, the cells were infected with SARS-CoV-2 at a multiplicity of infection (MOI) of 0.1 in the presence of either compound or dimethyl sulfoxide (DMSO) control. The dose–response curves were determined by quantification of CPE or by viral RNA copy numbers in the supernatant of infected cell at 72h post infection (p.i.). The cells and the cell culture supernatant would be saved at the time of CPE for additional analysis. According to an embodiment of the present invention, the test agent preparation comprises: • Four combination drugs are soluble in Dimethyl sulfoxide (DMSO). • All the four formulations would be tested in the range of 0-200 micromolar by taking properly spaced six concentrations. • Untreated cells would be considered as control. • No positive control is used in the study because there is no accepted positive control for SARS-Cov-19. • The interpretations would be largely based on the CPE value of the treatment profile.

According to an embodiment of the present invention, the process of assay protocol comprises of preparing 96 well plate with 80-90% confluent Vero cells in 200 µl medium. Next day, remove media in all wells, add fresh 200 µl medium containing drugs or DMSO control. After 2 hours of treatment, remove media in all wells and add 50 µl medium containing Virus and drugs or DMSO control. After, 2 hours of culture with virus, remove media in all wells and add 200 µl medium with FBS and drugs or DMSO control, incubate the plate in incubator. After 48 hours of culture (day 2), centrifuge the plate and collect carefully 75-100 µl culture supernatant for viral load estimation by RT-PCR and put the plate back to incubator. After 72 hours of culture (day 3), centrifuge the plate and collect culture supernatant and freeze for later use and cells will be used for estimating CPE. According to an embodiment of the present invention, the combination of therapeutic effective amount of nitazoxanide, albendazole and pyrantel are shown to be rapidly effective and produce faster resolution of SARS-CoV2 and fever than all other identified compounds. According to an embodiment of the present invention, the pharmaceutical composition comprising nitazoxanide along with albendazole and pyrantel were found to limit virus entry, reduces virus particles, inhibit both viral release and cell-to-cell transmission, and possess broad anti-viral activities which makes the purposing of existing drugs a timely and attractive alternative. According to an embodiment of the present invention, the pharmaceutical composition comprises at least one of pharmaceutically acceptable excipient. According to an embodiment of the present invention, the pharmaceutical composition comprises at least one of pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is selected from one or more of the group consisting of mannitol, sorbitol, a sorbic acid or potassium salt, sodium metabisulfite, sodium bisulfite, sodium thiosulfate, cysteine hydrochloride, thioglycolic acid, methionine, vitamin A, vitamin C, vitamin E, vitamin D, azone, disodium ethylenediaminetetraacetate, calcium sodium ethylenediaminetetraacetate, monovalent alkali metal carbonate, acetate, phosphate or an aqueous solution thereof, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, amino acid, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, a silicon derivative, cellulose and a derivative thereof, alginate, gelatine, polyvinylpyrrolidone, glycerol, propylene glycol, ethanol , Tween 60-80, Span-80, beeswax, lanolin, liquid paraffin, cetyl alcohol, a gallic acid ester, agar, triethanolamine, a basic amino acid, urea, allantoin, calcium carbonate, calcium bicarbonate, a surfactant, polyethylene glycol, cyclodextrin, 0-cyclodextrin, a phospholipid material, kaolin, talc, calcium stearate, magnesium, stearate, microcrystalline, poloxamer 407, poloxamer 188, phenethyl alcohol, methyl paraben, ethylparaben, benzalkonium bromide, sorbic acid, propylene glycol, polyethylene glycol 400,polyethylene glycol 2000, polyethylene glycol 4000, polyethylene glycol 6000, glycerine, mannitol, sorbitol, glucose, sodium chloride, and potassium chloride. According to an embodiment of the present invention, the pharmaceutical composition is a granule, tablet, capsule, oral, liquid, or mixture pellets. According to an embodiment of the present invention, the pharmaceutical composition can be administered as oral, parental, topical or any other mode of administration. According to an embodiment of the present invention, the combined preparation comprises of one or more dosage units of a nitazoxanide or its pharmaceutically acceptable salts thereof, an albendazole or its pharmaceutically acceptable salts thereof and a pyrantel or its pharmaceutically acceptable salts thereof. According to an embodiment of the present invention, the pharmaceutically acceptable excipients are selected from the group comprising diluents/fillers, binders, disintegrants, glidants, lubricants, solubilizers/surfactants/wetting agents, plasticizers, solvents, and the like. According to one embodiment of the present invention, the pharmaceutical compositions of the present invention can be in the form of modified release dosage form which include, but not limited to, controlled release, sustained release, extended release, prolonged release dosage form. Various rate controlling agents/techniques known in the art may be used to control the release of the drug from the dosage form. According to an embodiment of the present invention, the pharmaceutical composition may be in the form of tablets, capsules, tablets filled in capsule, mini- tablets filled in capsule, sachets containing powder or granules, pellets, and the like. The pharmaceutical composition is meant for once daily or twice daily administration. EXAMPLE OF PREPARATION OF NITAZOXANIDE, ALBENDAZOLE AND PYRANTEL FORMULATIONS The preparation of film coated tablets having some of the ingredients as listed below which can either include or exclude when prepared using conventional techniques:

Thus, the teachings of the present invention relates to the combination of nitazoxanide, albendazole and pyrantel administrations that improves the dosage regimen and combinational therapy that is not only effective and safe but also simple in administration resulting in a maximum of patients' compliance. Current therapy in the prior art has been found to be vulnerable if the strategy requires several drugs to be taken at different days and times. In addition, the rapid alleviation of symptoms due to the effect of several dosage often leads to abrupt end of the therapy as the patient feels well. This leads to high risk of recurrence of the disease and the development of severity due to an insufficient treatment. Compliance of the patient is, therefore, a crucial consideration and a key determinant in the successful treatment of COVID regarding both efficacy of the therapy and prevention of recurrence. Long duration regimens, complex dosing schemes, poor understanding of the patient of how or why to adhere to the recommended regimen as well as rapid perception of wellness induce the patient to discontinue the therapy prematurely. Therefore, it is a further need to provide the combinations which allow an anti- viral therapy that is not only effective, well-tolerated, prevents recrudescence, but also improves patients' compliance. The latter is achieved best if the treatment is simple and of short duration. According to an embodiment of the present invention, a combination of nitazoxanide, albendazole and pyrantel has been found that it provides a simple fixed-dose treatment administered one to two times a day for 5-10 days. This combination is unexpectedly well tolerated with a low rate of side-effects, and equally high cure-rates than in comparable treatments. Due to the simple dosing regimen the patients' compliance is excellent. As used herein the term “fixed-dose” means daily dosage of both antiviral that are same every day for a given treatment course, wherein the amount of the effective dose can be individually determined by the medical practitioner. According to an embodiment of the present invention, a method for the treatment of SARS-CoV2 in a 3-10 day course is also provided administering a therapeutically effective amount of the combination of nitazoxanide, albendazole and pyrantel is provided, wherein the dosage of nitazoxanide, albendazole and pyrantel for each day remains the same. The amount of the combination of nitazoxanide, albendazole and pyrantel required to be effective as an antiviral will, of course, vary and is ultimately at the discretion of the medical practitioner. The factors to be considered include the nature and severity of the SARS-CoV2 infection to be treated, route of administration, high or low transmission area, the immune status of the patient as well as age and weight of the patient. According to an embodiment, the present invention provides a pharmaceutical composition for treatment of SARS-CoV2 with existing pharmaceutical APIs and salts in novel composition, the respective raw materials, pharmaceutically acceptable carrier, and an extract can be prepared as granules, tablets, capsules, oral liquid, or mixture pellets. According to an embodiment of the present invention, the pharmaceutical composition can be a tablet, carrier as lactose or corn starch, magnesium stearate lubricants are added when required, mixed well, and then tableted. According to another embodiment of the present invention, when the present invention is made in capsules, the powder is filled in required doses in the capsule shells to form capsules. Since other modifications and changes varied to fit particular operating requirements and environments are apparent to those skilled in the art, the invention is not considered limited to the example chosen for purposes of disclosure and covers all changes and modifications which do not constitute departures from the true spirit and scope of this invention. The description of the present disclosure uses examples to disclose the invention, including the best mode, and also to enable any person skilled in the art to practice the invention, including making and using any devices or systems and performing any incorporated methods. The above are merely exemplary and illustrative of the inventive concept by, one of ordinary skill in the art in the specific embodiments described to make various modifications or additions, or a similar alternative manner, without departing from the invention. The invention, as defined in this disclosure or beyond the scope of the claims, should belong to the scope of the present invention. Further the dosage of the pharmaceutical composition in the present invention can carry out suitable variation according to the dosage form difference of administration object, route of administration or medicine, but take, guarantees that this pharmaceutical composition can reach effective blood drug level as prerequisite in mammalian body. While specific language has been used to describe the invention, any limitations arising on account of the same are not intended. As would be apparent to a person skilled in the art, various working modifications may be made to the method in order to implement the inventive concept as taught herein. The figures and the foregoing description give examples of embodiments. Those skilled in the art will appreciate that one or more of the described elements may well be combined into a single functional element. Alternatively, certain elements may be split into multiple functional elements. Elements from one embodiment may be added to another embodiment. For example, order of processes described herein may be changed and are not limited to the manner described herein. Moreover, the actions of any flow diagram need not be implemented in the order shown; nor do all of the acts need to be necessarily performed. Also, those acts that are not dependent on other acts may be performed in parallel with the other acts. The scope of embodiments is by no means limited by these specific examples.

Claims

WE CLAIM: 1. A pharmaceutical composition for treating SARS-CoV2 virus comprising a therapeutically effective amount of: a nitazoxanide or its pharmaceutically acceptable salts thereof; an albendazole or its pharmaceutically acceptable salts thereof; and a pyrantel or its pharmaceutically acceptable salts thereof.

2. The pharmaceutical composition as claimed in claim 1, wherein the pharmaceutical composition comprises the nitazoxanide in the ratio of 0.05% to 50% w/v.

3. The pharmaceutical composition as claimed in claim 1, wherein the pharmaceutical composition comprises the albendazole in the ratio of 0.02%to 30% w/v.

4. The pharmaceutical composition as claimed in claim 1, wherein the pharmaceutical composition comprises the pyrantel in the ratio of 0.0125 to 20% w/v.

5. The pharmaceutical composition as claimed in claim 1, wherein the pharmaceutical composition comprises at least one of pharmaceutically acceptable excipient.

6. The pharmaceutical composition as claimed in claim 1, wherein the pharmaceutical composition comprises at least one of pharmaceutically acceptable carrier.

7. The pharmaceutical composition as claimed in claim 1, wherein the pharmaceutical composition is a granule, tablet, capsule, oral, liquid, or mixture pellets.

8. The pharmaceutical composition as claimed in claim 1, wherein the pharmaceutical composition can be administered as oral, parental, topical or any other mode of administration.

9. The pharmaceutical composition as claimed in claim 1, wherein the pharmaceutical composition is also used in the treatment of Hepatitis B, Hepatitis C and Chikungunya virus.

10. A combined preparation comprising: one or more dosage units of a nitazoxanide or its pharmaceutically acceptable salts thereof; an albendazole or its pharmaceutically acceptable salts thereof; and a pyrantel or its pharmaceutically acceptable salts thereof.