# US 10,478,444 B2 · Methods and compositions for treating and preventing viral infections

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- Number: US 10,478,444 B2
- Document ID: US10478444B2
- Status: Granted
- Kind: B2
- Application number: 15/870,133
- Filed: 2018-01-12
- Published: 2018-05-17
- Granted: 2019-11-19
- Earliest claimed: 2009-10-27
- Assignees: Enterin, Inc.
- Applicants: Enterin, Inc.
- Inventors: Michael Zasloff
- Web page: https://patentgrep.com/patent/US10478444B2

## CPC

- A61K 31/575: substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
- A61K 31/56: Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K 38/212: {IFN-alpha}
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 31/12: Antivirals
- A61P 31/16: for influenza or rhinoviruses
- A61P 31/18: for HIV
- A61P 31/22: for herpes viruses
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

A method of treating or preventing a systemic viral infection in a mammal by administering a pharmaceutically acceptable composition selected from the group consisting of squalamine, an active isomer thereof, and an active analogue thereof, via a dosing regimen that delivers effective antiviral concentrations of squalamine. Also compositions for achieving the systemic antiviral effect.

## Claims

### Claim 1

An in vivo method of treating or preventing a viral infection in a mammal comprising administering a composition comprising:
(a) a pharmaceutically acceptable grade of squalamine or a pharmaceutically acceptable salt or derivative thereof in an amount sufficient to produce an antiviral effect, and
(b) at least one pharmaceutically acceptable carrier,
wherein the viral infection is caused by a virus selected from the group consisting of Yellow Fever, Eastern Equine Encephalitis virus, Human Immunodeficiency virus (HIV), “African Swine Fever Viruses,” Arbovirus, Adenoviridae, Arenaviridae, Arterivirus, Astroviridae, Baculoviridae, Bimaviridae, Bimaviridae, Bunyaviridae, Caliciviridae, Caulimoviridae, Circoviridae, Coronaviridae, Cystoviridae, EBV, Deltaviridae, Filviridae, Filoviridae, Flaviviridae, Iridoviridae, Mononegavirus, Myoviridae, Papiloma virus, Papovaviridae, Paramyxoviridae, Prions, Parvoviridae, Phycodnaviridae, Poxviridae, Potyviridae, Reoviridae, Retroviridae, Rhabdoviridae, Tectiviridae, Togaviridae, pox, papilloma, corona, influenza, sendai virus (SeV), sindbis virus (SINV), vaccinia viruses, West Nile, Hanta, viruses which cause the common cold, and any combination thereof, and
wherein the composition is systemically administered intravenously, subcutaneously, intramuscularly, orally, or by inhalation.

### Claim 2 (depends on 1)

The method of claim 1, wherein the effective daily dosing amount is about 0.1 to 20 mg/kg body weight.

### Claim 3 (depends on 1)

The method of claim 1, wherein the effective amount is administered in a regimen that achieves and maintains a tissue concentration of squalamine in body organs and tissues of between about 0.1 to about 200 μg/gram (tissue wet weight).

### Claim 4 (depends on 1)

The method of claim 1, wherein the mammal is human.

### Claim 5 (depends on 1)

The method of claim 1, wherein:
(a) the composition does not demonstrate an altered IC50 or IC90(drug concentration required to inhibit viral growth by 50% or 90% respectively) over time;
(b) the composition demonstrates an IC50 or IC90 which does not increase by more than 0%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, or 30% over time;
(c) the composition demonstrates an IC50 or IC90 which does not increase by an amount described in (b) over a time period selected from the group consisting of 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 1.5 months, 2 months, 2.5 months, 3 months, 3.5 months, 4 months, 4.5 months, 5 months, 5.5 months, 6 months, 6.5 months, 7 months, 7.5 months, 8 months, 8.5 months, 9 months, 9.5 months, 10 months, 10.5 months, 11 months, 11.5 months, 12 months, 1 year, 1.5 years, 2 years, 2.5 years, 3 years, 3.5 years, 4 years, 4.5 years, and 5 years; or
(d) any combination thereof.

### Claim 6

An in vivo method of treating or preventing a viral infection in a mammal comprising administering a composition comprising:
(a) a pharmaceutically acceptable grade of squalamine or a pharmaceutically acceptable salt or derivative thereof in an amount sufficient to produce an antiviral effect, and
(b) at least one pharmaceutically acceptable carrier,
wherein the viral infection is caused by a virus selected from the group consisting of:
(i) a Mononegavirus which is Paramyxoviridae, Morbillivirus, or Rhabdoviridae;
(ii) a Picomaviridae virus which is Rhinovirus or Poliovirus;
(iii) a Poxviridae virus which is Smallpox or Vaccinia;
(iv) a Reoviridae virus which is Rotavirus;
(v) a Retroviridae virus which is HTLV-I, HTLV-II, or Lentivirus;
(vi) a Togaviridae virus which is Rubivirus; and
(vii) any combination thereof.

### Claim 7

A method of treating or preventing a chronic disease suspected to be of viral origin, wherein the condition is selected from the group consisting of multiple sclerosis, Type I diabetes, Type II diabetes, atherosclerosis, cardiomyopathies, Kawaski disease, and aplastic anemia, and wherein the method comprises administering a composition comprising:
(a) a pharmaceutically acceptable grade of squalamine or a pharmaceutically acceptable salt or derivative thereof in an amount sufficient to produce an antiviral effect, and
(b) at least one pharmaceutically acceptable carrier.

### Claim 8 (depends on 1)

The method of claim 1, wherein the squalamine is administered in combination with at least one additional active agent to achieve either an additive or synergistic antiviral effect.

### Claim 9 (depends on 8)

The method of claim 8, wherein the additional active agent is administered via a method selected from the group consisting of
(a) concomitantly;
(b) as an admixture;
(c) separately and simultaneously or concurrently; and
(d) separately and sequentially.

### Claim 10 (depends on 8)

The method of claim claims 8, wherein the additional active agent is selected from the group consisting of:
(a) an antiretroviral agent;
(b) nucleoside or nucleotide reverse transcriptase inhibitors (NRTIs);
(c) non-nucleoside reverse transcriptase inhibitors (NNRTIs);
(d) nucleotide or nucleoside analogues;
(e) protease inhibitors (PIs);
(f) drugs based on “antisense” molecules;
(g) ribozyme antivirals;
(h) assembly inhibitors;
(i) release phase inhibitors;
(j) drugs which stimulate the immune system;
(k) fusion inhibitors/gp41 binders;
(l) fusion inhibitors/chemokine receptor antagonists;
(m) integrase inhibitors;
(n) hydroxyurea-like compounds;
(o) inhibitors of viral integrase;
(p) inhibitors of viral genome nuclear translocation;
(q) inhibitors of HIV entry;
(r) nucleocapsid zinc finger inhibitors;
(s) targets of HIV Tat and Rev;
(t) pharmacoenhancers;
(u) cytokines;
(v) lymphokines;
(w) an anti-inflammatory agent; and
(x) any combination thereof.

### Claim 11 (depends on 8)

The method of claim 8, wherein the additional active agent is selected from the group consisting of:
(a) an antiviral drug selected from the group consisting of Abacavir, Aciclovir, Acyclovir, Adefovir, Amantadine, Amprenavir, Arbidol, Atazanavir, Atripla, Boceprevir, Cidofovir, Combivir, Darunavir, Delavirdine, Didanosine, Docosanol, Edoxudine, Efavirenz, Emtricitabine, Enfuvirtide, Entecavir, Entry inhibitors, Famciclovir, Fomivirsen, Fosamprenavir, Foscarnet, Fosfonet, Fusion inhibitor, Ganciclovir, Ibacitabine, Imunovir, Idoxuridine, Imiquimod, Indinavir, Inosine, Integrase inhibitor, Interferon type III, Interferon type II, Interferon type I, Interferon, Lamivudine, Lopinavir, Loviride, Maraviroc, Molixan (NOV-205), Moroxydine, Nelfinavir, Nevirapine, Nexavir, Nucleoside analogues, Oseltamivir Peginterferon alfa-2a, Penciclovir, Peramivir, Pleconaril, Podophyllotoxin, Protease inhibitor, Raltegravir, Reverse transcriptase inhibitor, Ribavirin, Rifampicin, Rimantadine, Ritonavir, Saquinavir, Stavudine, Tenofovir, Tenofovir disoproxil, Tipranavir, Trifluridine, Trizivir, Tromantadine, Truvada, Valaciclovir, Valganciclovir, Vicriviroc, Vidarabine, Viramidine, Zalcitabine, Zanamivir, and Zidovudine;
(b) an NRTI selected from the group consisting of zidovudine/AZT, didanosinelddl, zalcitabine/ddC, stavudine/d4T, lamivudine/3TC, zidovudine/lamivudine, F-ddA, emtricitabine/FTC, dOTC (BCH-10652), Adefovir, Adefovir Dipivoxil; bis-POC PMPA, DAPD/DXG (active metabolite of DAPD); D-D4FC, GW420867, abacavir/159U89, CS-87 (3′azido-2′,3′-dideoxyuridine), and S-acyl-2-thioethyl (SATE)-bearing prodrug forms of beta-L-FD4C and P-L-FddC;
(c) an NNRTI selected from the group consisting of nevirapine, delavirdine, efavirenz, Emivirine/MKC442, AG-1549/S-1153, PNU-142721, DPC-961, DPC-963, GW420867X, CALANOLIDE A, and Propolis;
(d) a protease inhibitor selected from the group consisting of indinavir, ritonavir, saquinavir, nelfinavir, ABT378/r, BMS-232632 (an azapeptide), PNU-140690, PD-178390, BMS 232632 (an azapeptide), L-756,423 (an indinavir analog); DMP450 (a cyclic urea compound), AG-1776 (a peptidomimetic), VX-175/GW433908 (phosphate prodrug of amprenavir), CGP61755, and amprenavir;
(e) a fusion inhibitor/gp41 binder selected from the group consisting of T-20 (a peptide from residues 643-678 of the HIV gp41 transmembrane protein ectodomain) and T-1249);
(f) a fusion inhibitor/chemokine receptor antagonist selected from the group consisting of CXCR4 antagonists, AMD 3100 (a bicyclam), SDF-1, ALX404C (a cationic peptide), T22 (an 18 amino acid peptide), T134, T140, CCR5 antagonists, RANTES (9-68), AOP-RANTES, NNY-RANTES, TAK-779, CCR5/CXCR4 antagonists, NSC 651016 (a distamycin analog), CCR2B antagonists, CCR3 antagonists, CCR6 antagonists, MEP-lalpha, and MIP-1beta;
(g) an integrase inhibitor selected from the group consisting of dicaffeoylquinic (DFQA) acids; L-chicoric acid (a dicaffeoyltartaric (DCTA) acid), quinalizarin (QLC), AR 177, and naphthols;
(h) a hydroxyurea-like compound selected from the group consisting of BCX-34, ribonucleotide reductase inhibitors, inosine monophosphate dehydrogenase (IMPDH) inhibitors, VX-497, mycopholic acids, CellCept (mycophenolate mofetil);
(i) arylene bis(methylketone) compounds;
(j) inhibitors of HIV entry selected from the group consisting of AOP-RANTES, NNY-RANTES, RANTES-IgG fusion protein, soluble complexes of RANTES and glycosaminoglycans (GAG), and AMD-3100;
(k) nucleocapsid zinc finger inhibitors;
(l) pharmacoenhancers;
(m) cytokines and lymphokines selected from the group consisting of MIP-1alpha, MIP-1beta, SDF-1alpha, IL-2, aldesleukin/L2-7001, IL4, IL-10, IL-12, IL-13, interferons, IFN-alpha2a, IFN-alpha2b, IFN-beta, antagonists of TNFs, NFkappaB, GM-CSF, M-CSF, IL-10, agents that modulate immune activation, vaccines, gene-based therapies, antibodies; aryl hydrocarbon (AH) receptor agonists and antagonists, TCDD, 3,3′,4,4′,5-pentachlorobiphenyl, 3,3′,4,4′-tetrachlorobiphenyl, and alpha-naphthoflavone, antioxidants, gamma-L-glutamyl-L-cysteine ethyl ester (gamma-GCE);
(n) an anti-inflammatory agent selected from the group consisting of corticosteroids, nonsteroidal anti-inflammatory drugs, antihistamines, aminoarylcarboxylic acid derivatives, arylacetic acid derivatives, arylbutyric acid derivatives, arylcarboxylic acids, arylpropionic acid derivatives, pyrazoles, pyrazolones, salicylic acid derivatives, thiazinecarboxamides, e-acetamidocaproic acid, S-adenosylmethionine, 3-amino4-hydroxybutyric acid, amixetrine, bendazac, benzydamine, bucolome, difenpiramide, ditazol, emorfazone, guaiazulene, nabumetone, nimesulide, orgotein, oxaceprol, paranyline, perisoxal, pifoxime, proquazone, proxazole, and tenidap; and
(o) any combination thereof.

### Claim 12 (depends on 1)

The method of claim 1, wherein the method further comprises administration of an adjuvant.

### Claim 13 (depends on 12)

The method of claim 12, wherein the adjuvant is selected from the group consisting of cytokines and/or interleukins, alum, Lipid A, including monophosphoryl lipid A, bacterial products, endotoxins, cholesterol, fatty acids, aliphatic amines, paraffinic and vegetable oils, threonyl derivative, and muramyl dipeptide, alum, alum plus deoxycholate (ImmunoAg), MTP-PE (Biocine Corp.), QS21 (Genentech, Inc.), BCG, MPL, nonviable preparations of Corynebacterium, parvum, Monophosphoryl lipid immunomodulator, AdjuVax 100a, QS-21, QS-18, CRL1005, Aluminum salts, MF-59, and a Virosomal adjuvant.

### Claim 14 (depends on 1)

The method of claim 1, wherein the composition further comprises at least one antigen capable of eliciting an immune response.

### Claim 15 (depends on 14)

The method of claim 14, wherein the antigen is selected from the group consisting of viral and prion antigens.

### Claim 16 (depends on 13)

The method of claim 13, wherein the cytokine and/or interleukin is selected from the group consisting of IL2, IL3, IL4, IL5, IL6, IL7, IL8, IL-9, IL10, IL-11, IL12, IL13, IL-14, IL15, IIL16, IL-17, IL-18, IL-19, IL-20, IL-21, anti-CD40, CD40L, IFN-gamma, TNF-alpha, IL-Ialpha, IL-lbeta.

### Claim 17 (depends on 1)

The method of claim 1, wherein the composition is administered orally.

### Claim 18 (depends on 1)

The method of claim 1, wherein the composition is administered intravenously, subcutaneously, or intramuscularly.

### Claim 19 (depends on 1)

The method of claim 1, wherein the composition is administered by inhalation.

### Claim 20 (depends on 10)

The method of claim 10, wherein the drugs which stimulate the immune system are selected from the group consisting of interferons and synthetic antibodies.

### Claim 21 (depends on 11)

The method of claim 11, wherein the nucleocapsid zinc finger inhibitors are dithiane compounds.

### Claim 22 (depends on 11)

The method of claim 11, wherein the pharmacoenhancer is ABT-378.

### Claim 23 (depends on 11)

The method of claim 11, wherein the agents that modulate immune activation are selected from the group consisting of: cyclosporine and prednisone.

### Claim 24 (depends on 11)

The method of claim 11, wherein the vaccines are selected from the group consisting of: APL 400-003, recombinant gp120, recombinant gp120 fragments, bivalent (B/E) recombinant envelope glycoprotein, rgp120CM235, MN rgp120, SF-2 rgp120, gp120/soluble CD4 complex, Delta JR-FL protein, branched synthetic peptide derived from discontinuous gp120 C3/C4 domain, fusion-competent immunogens, Gag vaccines, Pol vaccines, Nef vaccines, and Tat vaccines.

### Claim 25 (depends on 11)

The method of claim 11, wherein the gene-based therapies are selected from the group consisting of: genetic suppressor elements (GSEs) and intrakines.

### Claim 26 (depends on 11)

The method of claim 11, wherein the antibodies are selected from the group consisting of: the anti-CXCR4 antibody 12G5, the anti-CCR5 antibody 2D7, the anti-CCR5 antibody 5C7, the anti-CCR5 antibody PA8, the anti-CCR5 antibody PA9, the anti-CCR5 antibody PA10, the anti-CCR5 antibody PA11, the anti-CCR5 antibody PA12, the anti-CCR5 antibody PA14, the anti-CD4 antibody Q4120, the anti-CD4 antibody RPA-T4, the anti-CCR3 antibody 7B11, the anti-gp120 antibody 17b, the anti-gp120 antibody 48d, the anti-gp120 antibody 447-52D, the anti-gp120 antibody 257-D, the anti-gp120 antibody 268-D, the anti-gp120 antibody 50.1, anti-Tat antibodies, anti-TNF-alpha antibodies, and the monoclonal antibody 33A.

### Claim 27 (depends on 6)

The method of claim 6, wherein the effective daily dosing amount is about 0.1 to 20 mg/kg body weight.

### Claim 28 (depends on 6)

The method of claim 6, wherein the effective amount is administered in a regimen that achieves and maintains a tissue concentration of squalamine in body organs and tissues of between about 0.1 to about 200 μg/gram (tissue wet weight).

### Claim 29 (depends on 6)

The method of claim 6, wherein the mammal is human.

### Claim 30 (depends on 6)

The method of claim 6, wherein:
(a) the composition does not demonstrate an altered IC50 or IC90 (drug concentration required to inhibit viral growth by 50% or 90% respectively) over time;
(b) the composition demonstrates an IC50 or IC90 which does not increase by more than 0%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, or 30% over time;
(c) the composition demonstrates an IC50 or IC90 which does not increase by an amount described in (b) over a time period selected from the group consisting of 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 1.5 months, 2 months, 2.5 months, 3 months, 3.5 months, 4 months, 4.5 months, 5 months, 5.5 months, 6 months, 6.5 months, 7 months, 7.5 months, 8 months, 8.5 months, 9 months, 9.5 months, 10 months, 10.5 months, 11 months, 11.5 months, 12 months, 1 year, 1.5 years, 2 years, 2.5 years, 3 years, 3.5 years, 4 years, 4.5 years, and 5 years; or
(d) any combination thereof.

### Claim 31 (depends on 6)

The method of claim 6, wherein the squalamine is administered in combination with at least one additional active agent to achieve either an additive or synergistic antiviral effect.

### Claim 32 (depends on 6)

The method of claim 6, wherein the method further comprises administration of an adjuvant.

### Claim 33 (depends on 6)

The method of claim 6, wherein the composition is administered orally.

### Claim 34 (depends on 6)

The method of claim 6, wherein the composition is administered intravenously, subcutaneously, or intramuscularly.

### Claim 35 (depends on 6)

The method of claim 6, wherein the composition is administered by inhalation.