Organosilicon carriers for use in treating infections and/or diseases caused by SARS viruses

Abstract

A method for treating, attenuating or inhibiting an infection and/or disease associated with a SARS virus in a subject is provided that includes administering a pharmaceutical composition to the subject. The composition includes an organosilicone carrier with one or more active substances that block and/or inhibit an ACE2 receptor in a host cell of the subject, the spike protein of a SARS virus and/or internal components of a virion of the SARS virus.

Claims

1. A method for treating, attenuating or inhibiting an infection and/or disease associated with a SARS CoV-2 virus in a subject, comprising administering a pharmaceutical composition to the subject, wherein the composition comprises three functional modules, each comprising an organosilicon carrier in the form of siosome nanoparticles, said modules comprising: Module 1: Siosomes with an ACE2 receptor inhibitor on the surface of the siosomes and/or an ACE2 inhibitor encapsulated, entrapped and/or conjugated in the siosomes, wherein said siosomes comprise at least one organosilicon compound covalently bound to a monosaccharide; Module 2: Siosomes with an inhibitor of the SARS virus spike protein on the surface of the siosomes and/or an inhibitor of the SARS virus spike protein encapsulated, entrapped and/or conjugated in the siosomes, wherein said siosomes comprise at least one organosilicon compound covalently bound to a monosaccharide; and Module 3: Siosomes with a polymerase inhibitor on the surface of the siosomes and/or a polymerase inhibitor encapsulated, entrapped and/or conjugated in the siosomes, wherein said siosomes comprise at least one organosilicon compound covalently bound to a cationic lipid and retaining a positive charge.

2. The method of claim 1, wherein the organosilicon carrier comprises at least one of sugar organosilicon or amino-sugar organosilicon compound according to the general formula (I): ##STR00024## whereby R.sub.1, R.sub.2, and R.sub.3 can be the same or different; whereby R.sub.1, R.sub.2, and R.sub.3 are selected from the group consisting of a monosaccharide, a disaccharide, an oligosaccharide, an amino sugar, a carbohydrate, and a nucleotide, wherein at least one of R.sub.1, R.sub.2, R.sub.3 is a monosaccharide; and whereby R4 is a fatty acid.

3. The method of claim 1, wherein the organosilicon carrier is in the form of a siosome that is capable of penetrating the SARS-CoV-2 virus and inhibiting its function.

4. The method of claim 1, further comprising one or more active substances, selected from the group consisting of monoclonal antibodies, carbohydrate, lipids, amino acids, peptides, proteins, nucleosides, inhibitors of the lung cell receptors, antiviral agents, antibacterial agents, genetic materials, antigens, antibodies, immuno-agents, anti-inflammatory agents, antitumor agents, cardio-protectors, hepato-protectors, GSH, oxidants, metal oxides, organosilicon compounds, Remdesivir, and corticosteroids, and wherein the one or more active substances are encapsulated, entrapped or conjugated in siosome nanoparticles.

5. The method according to claim 1, wherein the organosilicon carrier is obtained by: a) mixing at least one of organosilicon, sugar organosilicon, and amino-sugar organosilicon compounds in at least one of a solvent or vesicles, with the ACE2 receptor inhibitor, the inhibitor of SARS virus spike protein, and the polymerase inhibitor; b) homogenization, sonication and/or extrusion of the mixture, followed by c) separation of the free ACE2 receptor inhibitor, the inhibitor of SARS virus spike protein, and polymerase inhibitor d) sterile filtration of the mixture, e) lyophilization, and f) reconstitution to form a siosome of the multi-target and delivery system.

6. The method of claim 1, wherein the composition is a single or multiple dose formulation.

7. The method of claim 1, wherein the composition is administered via oral, rectal, vaginal, topical, nasal, intradermal, or parenteral administration, or as a transbuccal, sublingual, transmucosal or a sustained release formulation, wherein the parenteral administration is selected from the group consisting of subcutaneous, intravenous, intramuscular and infusion.

8. The method of claim 1, wherein the composition is administered in combination with additional antiviral therapies in patients with symptoms of a SARS-CoV-2 infection.

9. The method of claim 1, wherein the treatment comprises administration of a pharmaceutically effective dose of a second agent, selected from the group consisting of amino acids, carnitine/carnitine derivatives, neurotransmitters, vitamins, caffeine, antifibrotic agents, memory activating agents, neuroprotective agents, cardio-protective agents, antidiabetic agents, drugs for the prophylaxis and/or treatment of thrombosis, glutamate-antagonist, glutathione GSH, anti-Alzheimer's disease agents, antioxidants, anti-AIDS drugs, NSAIDS, antipsychotic drugs, buspirone, antidepressants, mood stabilizers, anticonvulsant, antigens, antibodies, genetic materials, catecholamines, hormones and sympatholytic adrenergic blocking agents.

10. The method of claim 1, wherein the at least one organosilicon compound covalently bound to a monosaccharide is at least one of 2-(Dim ethyldecylsilylethyl-b-D-glucopyranoside, 2-(Dimethyldodecylsilyl)ethyl-b-D-glucopyranoside, Butyldimethylsilyl-a-D-galactopyranoside, Dodecyldimethylsilyl-a-D-glucopyranoside, 1-O-Dioctadecylsilyl-di(2,3,4,6-O-tetraacetyl-b-D-galactopyranoside), 1-O-Dimethyl(dodecy])silyl-)2,3,4,6-O-tetraacetyl-b-D-glucopyranoside), 1-O-Dim ethyl(octadecyl)silyl-(2,3,4,6-O-tetraacetyl-b-D-glucopyranoside), Di(dodecanoyloxy)diphenylsilane, Dithexadecanoyloxy)diphenylsilane, or Di(undecanoyloxy)dimethylsilane.

11. The method of claim 1, wherein the organosilicon carrier further comprises neurotransmitters and/or amino acids as navigators incorporated on the surface of the siosomes, and/or conjugated to the siosomes, wherein said navigators are for the targeting of the brain and/or central nervous system.

12. The method of claim 1, wherein the organosilicon carrier further comprises gastro intestinal tract specific compounds, selected from the group consisting of N-acetyl cysteine, glutamine, fumarate, glycolic Acid, sodium glycol cholate, sodium deoxy cholate, sodium caproate, lectins, chitosan and Poly lactic acid, as navigators incorporated on the surface of the siosomes, and/or encapsulated, entrapped and/or conjugated in the siosomes, for the targeting of the gastro intestinal tract and/or colon.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1: Blocking the ACE 2 Receptors in the host cells using the ACE 2 inhibitor on the surface of the siosomes and the encapsulated ACE2 inhibitors in the siosomes after release.

(2) FIG. 2: Inhibition of the SARS-Co2 V: Blocking the spike (s) structural proteins with the effectors on the outer layer of the siosomes.

(3) FIG. 3: (1) Blocking the spike (s) structural protein (1) with the effectors on the outer layer of the siosomes (13).

(4) (2) Inhibition of the virus (RNA) using the different siosomes with encapsulated antivirus agent.

(5) FIG. 4: Overview: the multi-target and delivery organosilicon nanosystem (Siosomes®).

(6) FIG. 5: Structures of silanes/siosomes models with the different functions to provide a novel multi-target and delivery organosilicon siosomes nanosystem.

(7) FIG. 6: Specific targeting of the GI using the Siosomes as multi-target and delivery system.

(8) FIG. 7: Multi-target and delivery system using the Siosomes: Multi-Lameller siosomes vesicles with active drug.

(9) FIG. 8: Drug Delivery using Siosomes—Use of multi-lameller Siosomes vesicles as multi-target and delivery which reach the site of action with at least one intact vesicle layer and still contain the prodrug polymer complex.

(10) FIG. 9: Drug delivery and targeting system using the multi-target and delivery system the Siosomes-Flexibility of design for the delivery of lipid, genetic materials, proteoms, antigens and antibiotics.

(11) FIG. 10: Effect against influenza virus A (H3N2)—Example for the antiviral activity of SIL 25.

(12) FIG. 11: Importance of sugar residues in the sugar Silanes and Siosomes for targeted binding against viruses, bacteria and cancer cells (Salama).

(13) FIG. 12: The Siosomes-Drug conjugation (SDC) and Antibody Siosomes conjugates (ASC)—Using Linkers (cleavable/non-cleavable).

(14) FIG. 13: The Siosomes-Genetic Material-Conjugate (SGMC and Siosomes-Antiviral Drugs-Conjugates (SADC)-Using Linkers (cleavable/non-cleavable).

(15) FIG. 14: Flowchart for the preparation of the Multi-Target and delivery siosomes system.

(16) FIG. 15: Preparation of the combined multi-target and delivery siosomes for the attenuation, prevention and/or treatment of the infection caused by the SARS Co 2V.

EXAMPLES

(17) The examples provided herein represent practical support for particular embodiments of the invention and are not intended to limit the scope of the invention. The examples are covering the following areas on the Multi-Target- and delivery Structure and effects of the siosomes and the compounds of the invention for the prophylaxis, prevention, attenuation, and/or therapy of said viral inflammations, Alzheimer's, neurodegenerative diseases, and neuromuscular degenerative diseases.

Example 1

(18) Preparation of the Multi-Target and Delivery Siosomes for the Attenuation, Prevention and or Treatment of the Inflammations Caused by the SARS Co 2-V

(19) It is well recognized in the medical field that the most effective procedures for treating localized diseases and/or specific targeting of an organ, cell, receptors or cell compartments is to direct a pharmaceutical agent or active substance to the affected area, thereby avoiding undesirable toxic effects of systematic treatment.

(20) Techniques and methods currently used in this invention to deliver active substances to specific target sites within the body involve the utilization of time-release delivery systems and the appropriate design of the multi-target and delivery system as mentioned below.

(21) To reach the site of action, the API has to cross many biological barriers, such as other organs, cells and intracellular compartments, where it can be inactivated or express undesirable effects on organs and tissues that are not involved in the pathological process.

(22) To achieve according to the invention the required objectives of the specific multi-target and delivery systems defined as:

(23) 1. Function of the Module 1-3

(24) Module 1 to block and/or inhibit the ACE2 receptors in the host cells.

(25) Module 2 to block and/or inhibit the spike protein of the SARS Co2 V virus.

(26) Module 3 to block, interact, change and/or inhibit the internal compartments of the virion in the SARS Co2 V (e.g. RNA-protein nucleocapsid, membrane protein).

(27) Module 3.1: “Inhibition of the SARS-CoV-2 virus” (Inhibition of viral RNA dependent RNA polymerase, blocking late stage of virus assembly) using in siosomes encapsulated, entrapped and/or conjugated polymerase inhibitors and siosomes transfection agents.

(28) Module 3.2: “Inhibition of the SARS-CoV-2 virus” (Inhibition of viral RNA dependent RNA polymerase, blocking late stage of virus assembly) using polymerase inhibitors on the surface (outer layer) of the siosomes connected via cleavable and/or non-cleavable LINKERS.

(29) 2. Composition of the Module 1-3

(30) According to the invention the method is characterized that the modules have the compositions:

(31) Module 1: Siosomes with specific ACE2 receptor inhibitor and carbohydrate molecule on the surface of the siosomes and/or specific ACE2 inhibitor encapsulated, entrapped and/or conjugated in the siosomes.

(32) Module 2: Siosomes with specific inhibitor of the spike protein and carbohydrate molecule on the surface of the siosomes.

(33) Module 3.1: Siosomes encapsulated, entrapped and/or conjugated polymerase inhibitors and siosomes transfection agents.

(34) Module 3.2 Siosomes with polymerase inhibitors on the surface (outer layer) of the siosomes connected via cleavable and/or noncleavable LINKERS.

(35) Examples for structures of silanes and sugar silanes to provide a novel multi-target and delivery organosilicon siosomes nano system

Example 1.1 (FIG. 1)

(36) MODULE 1: Blocking the ACE2 Receptors e.g. in the human lung epithelial cells, heart cell, Kidney, GI using the siosomes nano carrier system. Each of the following 10 silanes/siosomes represent ONE module 1.

(37) The 10 experiments in Example 1 with the ACE 2 inhibitor (captopril) will provide the following comparative data on the inhibition of the ACE2 receptor using the different compositions defined in the table. The objective will be to assess the composition which will achieve the best targeting and efficacy. (Figure)

(38) 1. Encapsulation of captopril in the siosomes

(39) 2. One molecule Captopril on the surface of the siosomes

(40) 3. Captopril on the surface and encapsulated in the siosomes

(41) 4. Two molecules of captopril on the surface of the siosomes

(42) 5. Influence of the sugar molecules on the surface of the siosomes One molecule sugar Two molecules of sugar Three molecules of sugar

(43) 6. Influence of amino acids on the surface of the siosomes

(44) 7. Influence of navigators on the surface One molecule of navigator (amino acid) Two molecules of navigator (amino acid)

(45) TABLE-US-00001 TABLE 1 Silanes/Siosomes MODULE 1 Compounds encapsulated, Silane/Siosomes Compounds on the Compounds in entrapped and/or No. surface/outer layer the mono/bilayer conjugated in the MODULE 1 of the siosomes of the Siosomes siosomes 1 R1: Monosaccharide R3: Lipid chain ACE2-Receptor R2: ACE2- Receptor R4: Lipid chain Inhibitor Inhibitor 2 R1: Monosaccharide R3: Lipid chain ACE2-Receptor- R2: Monosaccharide R4: Lipid chain Inhibitor 3 R1: Monosaccharide R4: Lipid Monosccharide R2: ACE2 Receptor Inhibitor R3: ACE2-Receptor Inhibitor 4 R1: Monosaccharide R4: Lipid ACE2 Receptor R2: Monosaccharide Inhibitor R3: Monosaccharide 5 R1: Monosaccharide R4: Lipid ACE2-Receptor R2: ACE2-Receptor Inhibitor Inhibitor R3: Amino Acid (Leucin) 6 R1: Monosaccharide R4: Lipid ACE2-Receptor R2: ACE2-Receptor Inhibitor Inhibitor R3: Navigatiting molecule 7 R1: Monosaccharide R4: Lipid ACE2-Receptor R2: Monosaccharide Inhibitor R3: Navigating molecule 8 R1: Monosaccharide R3: Lipid ACE2-Receptor R2: Navigating molecule R4: Lipid Inhibitor 9 R1: Monosaccharide R4: Lipid ACE2-Receptor R2: Navigating molecule Inhibitor R3: Navigating molecule 10 R1: Monosaccharide R3: Lipid ACE2-Receptor R2: Amino Acid R4: Lipid Inhibitor

Example 1.2: (FIG. 2)

(46) MODULE 2: Blocking the spike glycoprotein (S protein) on the virion surface with its subunits S1 and S2 of the SARS-CoV-2 Virus using the siosomes nanocarrier system. Each of the following 4 silanes/siosomes represent ONE module 2

(47) Module 2 (FIG. 2)

(48) 1. Influence of monosaccharide (responsible for cell recognition and communication) on the surface of the siosomes on blocking of the spike glycoprotein (s protein): The objective will be to investigate the interaction of the monosaccharide with the spike glycoprotein and the blocking and inhibition of its targeting to the ACE2 receptors. One molecule sugar Two molecules sugar Three molecules sugar 2. Influence of amino acids and sugar on the surface of the siosomes. This to understand the impact of amino acids on the modification of the spike glycoprotein. 3. Influence of monosaccharide, dipeptides, polypeptides encapsulated in the siosomes.

(49) TABLE-US-00002 TABLE 2 Silanes/Siosomes MODULE 2 Compounds encapsulated, Silane/Siosomes Compounds on the Compounds in entrapped and/or No. surface/outer layer the mono/bilayer conjugated in the MODULE 2 of the siosomes of the Siosomes siosomes 1. R1: Monosaccharide R3: Lipid Monosaccharide R2: Monosaccharide R4: Lipid 2. R1: Monosaccharide R3: Lipid Polysaccharide R2: Monosaccharide R4: Lipid 3. R1: Monosaccharide R4: Lipid Monosaccharide R2: Monosaccharide R3: Monosaccharide 4. R1: Monosaccharide R4: Lipid Dipeptide/polypeptide R2: Monosaccharide R3: Amino Acid 5. R1: Monosaccaride R3: Peptid Dipeptide/polypeptide R2: Monosaccharide R4: Lipid

Example 1.3.1 (FIG. 3)

(50) MODULE 3.1: “Inhibition of the SARS-CoV-2 virus” (Inhibition of viral RNA dependent RNA polymerase, blocking late stage of virus assembly) using in siosomes encapsulated, entrapped and/or conjugated polymerase inhibitors. Each of the following 8 silanes/siosomes represent ONE module 3.1
Module 3.1 (FIG. 3) 1. Influence of monosaccharide on the surface of the siosomes on the transfection of the siosomes into the virus and the inhibition of the virion: The objective will be to investigate the impact of the monosaccharide on the transfection of the siosomes and the inhibition of the virion One molecule sugar Two molecules sugar Three molecules sugar 2. The influence of the different polymerase inhibitors which are encapsulated in the siosomes. The objective will be to investigate the influence of the sugar molecules on the targeting and efficacy of the polymerase inhibitors. Camostat R Remdisivir Kaleta R Avigan MRNA Sodium meta arsenite

(51) TABLE-US-00003 TABLE 3.1 Silanes/Siosomes MODULE 3.1 Compounds encapsulated, Silane/Siosomes Compounds on the Compounds in entrapped and/or No. surface/outer layer the mono/bilayer conjugated in MODULE 3.1 of the siosomes of the Siosomes the siosomes 1 R1: Monosaccharide R3: Lipid Polymerase inhibitor R2: Monosaccharide R4: Lipid “Camostat mesylate” 2 R1: Monosaccharide R4: Lipid Polymerase Inhibitor R2: Monosaccharid “Camostat Mesylate” R3: Monosaccharide 3 R1: Monosaccharide R3: Lipid m-RNA R2: Monosaccharide R4: Lipid linked with PEG 4 R1: Monosaccharide R3: Lipid Polymerase inhibitor: R2: Monosaccharide R4: Lipid Remdisivir 5 R1: Monosaccharide R4: Lipid Polymerase inhibitor: R2: Monosaccharid Remdisivir R3: Monosaccharide 6 R1: Monosaccharide R3: Lipid sodium metaarsenite - R2: Monosaccharide R4: Lipid tolemerase inhibitor 7 R1: Monosaccharide R3: Lipid Protease inhibitor: R2: Monosaccharide R4: Lipid Kaletra 8 R1: Monosaccharide R3: Lipid AVIGAN (Favipiravirand R2: Monosaccharide R4: Lipid metabolite Favipiravir- ribofuranosyl-5′- triphosphate(Favipiravir- RTP): Inhibitor of viral RNA dependent RNA Polymerase

Example 1.3.2 (FIG. 3)

(52) MODULE 3.2 “Inhibition of the SARS-CoV-2 virus” (Inhibition of viral RNA dependent RNA polymerase, blocking late stage of virus assembly) using polymerase inhibitors on the surface (outer layer) of the siosomes connected via cleavable and/or noncleavable LINKERS and cationic siosomes as “transfection agents”. Each of the following 4 silanes/siosomes represent ONE module 3.2 The objective will be to investigate the following; 1. Influence of the polymerase inhibitor on the surface (outer layer) of the siosomes and cationic siosomes as “transfection agents” on the inhibition of the virion. 2. The influence of the polymerase inhibitors Camostat mesylate Remdisivir

(53) TABLE-US-00004 TABLE 3.2 Silanes/Siosomes MODULE 3.2 Compounds on the Compounds surface/outer layer encapsulated, Silane/Siosomes of the siosomes Compounds in entrapped and/or No. Linker with the the mono/bilayer conjugated in MODUL 3.2 Siosomes of the Siosomes the siosomes. 1. R1: Monosaccharide R3: Lipid Polymerase inhibitor R2: Camostat-Linker- R4: lipid “Camostat mesylate Siosomes 2. R1: Monosaccharide R3: Lipid Polymerase inhibitor: R2: Remdisivir - R4: Lipid Remdisivir Linker-Siosomes 3. R1: Monosaccharide R3: Lipid Protease inhibitor: R2: Kaletra -Linker- R4: Lipid Kaletra Siosomes 4. R1: Monosaccharide R3: Lipid AVIGAN R2: Monosaccharide R4: Avigan- Linker-Siosomes

Example 2

(54) Determination of the cytotoxicity and antiviral Activity of 43 selected silanes and sugar silane/siosomes with different chemical groups and structures

(55) I. Outlines

(56) (1) Screening of the 43 silanes and sugar silanes for antiviral activity against model viruses belonging to taxonomic groups including causative agents of infections in which applications of chemotherapy is strongly indicated. a) Enterovirus B (or C) (Family Picornaviridae) b) Bovine Viral Diarrhea virus (a surrogate hepatitis C virus) (Family Flaviviridae) c) Influenza virus A (Family Orthomyxoviridae) d) Respiratory syncytial virus (Family Paramyxoviridae) e) Human adenovirus 2 (or 5) (Family Adenoviridae) f) Herpes simplex virus type 1, and g) Human cytomegalovirus (Family Herpesviridae) h) Vaccinia virus (Family Poxviridae) (2) The antiviral screening was performed in-vitro in cell cultures. The CPE inhibition test in monolayer cell cultures (in micro plates); photometrical (optical density) measurement of neutral red uptake was carried out. In parallel, cytotoxicity has been determined. Compounds manifesting antiviral effects has been included in some additional tests for in-vitro antiviral activity illustration.
Determination of Cytotoxicity

(57) The neutral red uptake assay based on the initial protocol described by Borenfreund and Puerner (1984) was used. Monolayer cells in 96-well plates are inoculated with 0.1 ml of the tested solution in several serial dilutions performed in a maintenance medium. Cells inoculated with 0.1 ml maintenance medium (no compound in the medium), serve as a control. Each tested dilution is inoculated in 6 wells of the cell culture plate. Then, the cells were incubated at 37° C. in a humidified atmosphere with 5% CO.sub.2 and the cell vitality at the 48′, 72.sup.nd or 96.sup.th hour was estimated (following light microscopy observation) using ELISA reader at OD.sub.540 nm. The 50% cytotoxic concentration (CC.sub.50) is calculated in comparison to the cell control by applying the regression analysis with the help of Origin 6.1 computer program.

(58) Determination of Antiviral Activity

(59) The cytopathic effect (CPE) inhibition test is used for measuring the antiviral effect. Monolayer cells in 96-well plates are inoculated with 0.1 ml virus suspension containing 100 CCID.sub.50. After an hour for virus adsorption (two hours in the case of HRSV-A2) in a humidified atmosphere at 37° C. and 5% CO.sub.2, excessive virus is discarded and cells are inoculated with 0.2 mL of maintenance medium containing serial 0.5 lg dilutions of the tested preparation. Mock-infected cells are left for cell and toxicity controls. The virus CPE is scored daily by inverted light microscope (Olympus CK40, Japan) at 125× and 400× magnification on a 0-4 basis (4 representing total cell destruction) till the appearance of its maximum in the virus control wells (with no compound in the maintenance medium)—the 48′ hour p.i. for PV1, 72.sup.nd hour for HRSV-A2 and the 4th day (96.sup.th hour) p.i. for HAdV-2. When maximum CPE in the virus control wells is reached, cells are processed according to the neutral red procedure described above. The percent of virus CPE protection is calculated by the following formula [Pannecouque et al., 2008]:

(60) $\frac{{\mathrm{meanOD}}_{\mathrm{Test}}-{\mathrm{meanOD}}_{\mathrm{VC}}}{{\mathrm{meanOD}}_{\mathrm{TC}}-{\mathrm{meanOD}}_{\mathrm{VC}}}\times 100$
(OD.sub.Test−OD.sub.VC)/(OD.sub.TC−OD.sub.VC)×100(%), where OD.sub.Test is the mean optical density (OD) of the test sample, OD.sub.VC—the absorbance of the virus-infected control (no compound in the maintenance medium), and OD.sub.TC—the OD of the mock-infected control (toxicity control).

(61) The 50% virus inhibitory concentration (IC.sub.50) is determined by applying the regression analysis with the help of Origin 6.1 computer program and it is expressed as the concentration that achieves 50% protection of virus-infected cells.

(62) The selectivity index (SI) is evaluated as the ratio between CC.sub.50 and IC.sub.50 (SI=CC.sub.50/IC.sub.50).

(63) Each of the tests described above was done in triplicate to quadruplicate, with four cell culture wells per test sample.

(64) Antiviral Activity

(65) The screening carried out for antiviral activity in vitro (in cell culture experiments) of 43 Silanes and sugar silanes according to General Formel 1 embraced eight viruses belonging to taxonomic entities (families) including causative agents of infections to which chemotherapy is indicated. The results obtained demonstrated a marked activity of Silane No. 27 (SIL27) only against human cytomegalovirus: SI=30.9. Silane No. 20 (SIL 20) showed a marked activity as well toward the cytomegalovirus: at a low m.o.i. (3.2 CCID50 per microplate well) Close to borderline effect against this virus cytomegalovirus was found by Silanes SIL2, SIL7, SIL15, SIL 19 and SIL 34 and Silanes SIL 2, SIL3 and SIL 25 showed marked activity toward influenza virus A(H3N2). Silane SIL 9 showed marked activity versus vaccinia virus. No one of the Silanes manifested activity towards PV1, BVDV, RSV, HuAdV2 and HSV type 1.
Cytotoxicity Activity

(66) As concerns the cytotoxicity it was established a strong variation towards different cell Cultures used.

(67) Higher cytotoxicity values (CC50<20 μM) were recorded as follows: Silanes 1, 2, 5, 21, 22, 23, 24, 26 and 36 towards HEp-2 cells, Silanes 1, 2, 3, 5 and 25—toward CT cells, Silanes 2, 3 and 26 toward MDCK cells, Silanes 1, 2 and 3 toward MDBK cells, Silanes 4, 7, 22, 23, 24 and 32 toward MRC-5 cells Silanes 5, 21 and 36 vs Vero cells.

(68) Summarizing the cytotoxicity data it could indicate several compounds possessing wider toxicity, on more than one cell culture: Silane 1—on HEp-2 cells, CT cells and MDBK cells Silane 2—on HEp-2, CT and MDCK cells Silane 5—on HEp-2, CT, MDBK and Vero cells Silanes 22, 23 and 24—on HEp-2 and MRC-5 cells Silane 26—on HEp-2 and MDCK cells Silane 36—on HEp-2 and Vero cells. Silane 21 manifested a marked cytotoxicity only on HEp-2 cells. Comparing the cytotoxicity susceptibility of the different cell cultures species it could marked the higher susceptibility of the HEp-2 cells.

(69) Evidently, the realization of quantitative structure-activity relationship (QSAR) of the silanes included in this study would contribute for the further planned synthesis of active antiviral compounds, especially directed against HCMV.

(70) Discussion

(71) Antiviral Activities

(72) Surprisingly and unexpected 10 silanes and sugar silanes have shown antiviral activities with the following investigated enveloped viruses (Table 4):

(73) Human Cytomegalvirus (HCMV) consists of an outer lipid bilayer envelope, composed of various viral glycoproteins.

(74) Influenza virus A (virion as the infectious particle) is roughly spherical. It is an enveloped virus—that, the outer layer of the viron is a lipid membrane.

(75) Vaccinia virus (VACV or VV) is a large, complex, enveloped virus.

(76) These viruses are enveloped viruses as the SARS viruses and the SARS Co-2V.

(77) The investigated silanes and sugar silanes have different chemical structures such as the following differences: Different residues on the surface of the siosomes (sugar molecules, Phenyl, methyl . . . ) Number and length of the lipid chain(s) Electrical charges of the molecule Physicochemical properties (solubility, stability, chemical reactions) The 7 Silanes 2, 3, 7, 9, 15, 19, 20 belong to the “Sugar Silanes” with sugar molecules on the surface of the siosomes nano particles. The 3 Silanes 25, 27, 34 belong to the Silanes and have other molecules than sugar on the surface of the siosomes.

(78) With reference to the unexpected antiviral results in the invention, it be concluded: The sugar silanes and silanes are suitable as multi-target and delivery systems The sugar molecules on the surface of the siosomes are very relevant for the interaction with the receptors in the host cell, and/or they block it. The sugar and other residues on the surface of the siosomes are relevant for the recognition, communication, interaction, blocking and/or inhibition of the virus. The siosomes could penetrate and transfect the envelope of the virus and inhibit the virion activities. The siosomes are appropriate for the encapsulation, entrapment, and/or conjugation of the different active substance including, but not limited to antiviral agents, antibacterial, transfection agents, peptides and oxidants. The siosomes are appropriate to be as inhibitors to the viruses via fusion with the Virus. The silanes and sugar Silanes are appropriate according to the invention to be linked to antibodies, and active agents. The use of antiviral silanes and sugar silanes could have additive antiviral activities when it is encapsulated, entrapped or conjugated with antiviral agents.
Cytotoxicity Activities

(79) Surprisingly and unexpected that a number of the investigated silanes and sugar silanes with different molecular structure have shown cytotoxicity. As a total 14 Silanes and Sugar Silanes have shown cytotoxicity activities. They are as follows: The 6 silanes 1, 2, 3, 4, 5, 7, belong to the “Sugar Silanes” with sugar molecules on the surface of the siosomes nano particles. The 8 Silanes 21, 22, 23, 24, 25, 26, 32, 36 belong to the silanes and have other molecules than sugar on the surface of the siosomes. It is also unexpected that only the following three (3) sugar silanes 2, 3, 7 and one (1) non-sugar silane No. 25 have shown in addition to the antiviral activities cytoxicity.

(80) It is surprisingly unexpected that 23 Silanes and Sugar Silanes of the 43 investigated compounds approximately 53% are inert. The silanes and Sugar silanes and their derivatives according to the invention provide a great diversity of multi-target and delivery carrier for the encapsulation, entrapment, conjugation with active substances as for the attenuation, prevention and/or treatment of viral infections and diseases caused by SARS viruses such as SARS Co 2-V.

(81) TABLE-US-00005 TABLE 4 Silanes and sugar silanes with anti-virus activities Molecular MW No. Structure and Name Formula (Da) Sil 2 C20H42O6Si 406.64 Sil 3 C22H46O6Si 434.69 Sil 7 C12H26O6Si 294.42 Sil 9 C20H42O6Si 406.64 Sil 15 0 C64H112O20Si 1229.68  Sil 19 C28H50O10Si 574.79 Sil 20 C34H62O10Si 658.95 Sil 25 C36H56O4Si 580.93 Sil 27 C44H72O4Si 693.15 Sil 34 C24H48O4Si 428.73

(82) TABLE-US-00006 TABLE 5 Cytotoxicity and antiviral activity of silanes on the replication of influenza A(H3N2) Aichi virus in MDCK cells (as an example) Compound CC.sub.50 (μM) IC.sub.50 (μM) SI Sil 1 30.0 — — Sil 2 18.0 7.38 2.43 Sil 3 19.2 6.6  2.9  Sil 5 32.0 — — Sil 7 444.2 — — Sil 8 23.5 — — Sil 9 444.2 — — Sil 10 156.0 — — Sil 11 444.2 — — Sil 19 65.5 — — Sil 20 2533.0 — — Sil 21 21.0 — — Sil 22 42.9 — — Sil 23 93.2 — — Sil 24 32.0 — — Sil 25 55.9 9.6  5.82 Sil 26 19.0 — — Sil 27 317.0 — — Sil 31 52.1 — — Sil 32 45.2 — — Sil 33 155.0 — — Sil 34 2533.0 — — Sil 42 398.0 — — Sil 43 137.0 — — Sil 44 537.0 — — Sil 45 560.0 — — Rimantadine 94.0 0.03 3133.0  
Effect Against Influenza Virus a (H3N2)
FIG. 10: Cytotoxicity and antiviral activity of silanes on the replication of influenza virus A/Aichi/2/68 (H3N2) in MDCK cells—SIL 25

Example: Sugar Silane—SIL 25

Example 3

(83) Tissue Distribution Results for Study Number IPSS E010 after Repeated (Seven Continuous Days 1 mg Each Day) Administration of Cis-Oxoplatin (Oral and Intravenous) and Cis-Oxoplatin Siosome Complex (Intravenous)—No Non Treatment Period (Rats Sacrificed 24 Hrs after Last Administration)
Objectives

(84) To investigate the influence of the encapsulation of cis-oxoplatin as a reference compound from the active substances according to the invention on the tissue distribution in comparison to the un-encapsulated cis-oxoplatin.

(85) The molar adjustment had been taken into consideration for the determination of the distributed cis-oxoplatin in the different tissues.

(86) The cis-oxoplatin siosome complex has the highest concentration in blood, kidney and adipose tissue.

(87) Standard cis-oxoplatin administered intravenously has the highest platinum concentration in the stomach and the liver.

(88) The standard cis-oxoplatin oral has the highest accumulation of platinum in the spleen and lungs.

(89) The highest tissue accumulation for all treatment groups was found in the kidneys, though the blood had the highest concentrations.

(90) TABLE-US-00007 TABLE 6 Percentage of Total Platinum in Each Tissue as a Percentage of Administered Total Platinum Cis-Oxoplatin Standard Cis- Standard Cis- Siosome Complex Tissue Oxoplatin Oral Oxoplatin i.v. i.v. Stomach 0.011% 0.022% 0.026% Liver 0.201% 1.190% 1.032% Lung 0.048% 0.039% 0.054% Kidney 0.170% 0.577% 0.789% Spleen 0.035% 0.045% 0.054% Adipose Tissue 0.005% 0.035% 0.075% Blood 0.306% 0.658% 0.835% Total 0.776% 2.566% 2.865%

(91) Surprisingly and unexpected that the tissue with the highest actual content of total platinum by mass was the liver for all treatment groups. The total measured mass as a percentage of the platinum initially administered within the tissues and blood collected of total platinum was 0.776% for the standard cis-oxoplatin administered orally, 2.566% for the standard cis-oxoplatin administered intravenously and 2.865% for the cis-oxoplatin encapsulated in siosomes administered intravenously. These values are percentage of total platinum found in the six organs tested compared to the amount of total

(92) Discussion of the Results

(93) In this example for the tissue Distribution study in rats. Cis-oxoplatin as ant-viral agent encapsulated in the siosomes and the animals have been administered the two preparations intravenously.

(94) Cis-oxoplatin has been encapsulated in the siosomes and Free cis-oxoplatin as (un-encapsulated). Both products were administered intravenously to the animals.

(95) The unexpected and surprisingly results have shown the following: The concentration and content of cis-oxoplatin in the following organs and blood for the encapsulated cis oxoplatin formulation were higher compared to the un-encapsulated cis-oxoplatin: Lung: 38% Kidneys: 36% Spleen: 20% Blood: 27% Blood and the organs lungs, kidneys and spleen and especially the lungs are the most important organs along with the heart and liver for the infection activities and organ dysfunction caused by the virus and particularly SARS Co2V and the other SARS viruses.

(96) It is therefore a great advantage of the encapsulated and/or entrapped antiviral agents of the active substances. This will increase the accumulation of the antivirus agent in these relevant organs and tissues which will be infected by the virus. This will improve in addition the bioavailabilty of the antiviral drug in the blood and organs and increase the efficacy.

(97) Furthermore In there is the possibility to design and customize further multi-target and delivery siosomes in order to increase these positive antiviral effects.

Example 4

(98) Synthesis of a Sugar Silane for the Conjugation to Monoclonal Antibodies and Protein for the Preparation of the Siosomes-Monoclonal Antibody Conjugate

(99) Description

(100) Synthesis of a sugar silane for use for conjugation to monoclonal antibodies and proteins for the preparation of Siosomes. The sugar silane will be the SIL 17 (with 2 sugar molecules replaced with: (1) Tri-peptide QPG (Si-QPG) (2) Non-cleavable linker contains a spacer and a maleimide functional group. The spacer could be with short chain. This means no need for PEG to avoid steric hindrance for the reaction of the maleimide with the mAB.
Mesylate-Azide Route

(101) This approach is based on partial conversion of a di-mesylate in a mesylate azide intermediate. A Grignard addition of dodecyl-magnesium chloride to dodecyl-trichlorosilane followed by a double Grignard with vinyl magnesium chloride to give intermediate 3. Hydroboration and treatment with MsCl should give dimesylate 5. Treatment with a single equivalent of azide will hopefully result in mixture containing compound 6 as isolatable component. The yield is expected to be low because a literature reference used a similar approach and isolated the mono-mesylate in 18% yield. The mesylate may be used to alkylate the tripeptide. Subsequent reduction of the azide will give amine 9 that can be coupled to active ester 10.

(102) ##STR00016## ##STR00017##

(103) This is one of the many possibilities that the silanes, sugar silanes and their derivatives offer to conjugate with the monoclonal antibodies, and proteins to produce siosomes with the monoclonal antibodies on the surface SABC (Siosomes Antibody Conjugate) (FIG. 12).

(104) In addition, as further possibilities are: the production of the following siosomes conjugates: Siosomes-Genetic materials conjugate SGMC (FIG. 13) Siosome Antivirus Drugs Conjugates SADC (FIG. 13) Siosomes-Corticosteroids Conjugates SCC (Fig) Siosomes-Active substances-conjugates such as polymers, etc.

(105) It is unexpected that the manufacture of Siosome Conjugates with pharmaceuticals such as anti-cancer, antivirus, antibiotics and corticosteroids will have many advantages compared to the process known in the prior art as “ADC” Antibody Drug Conjugate. A number of the advantages of the siosomes are: Easy chemistry—chemical reactions between the silane molecules or sugar silanes and the antivirus, antibiotics, non-cytotoxic compounds instead of reaction/conjugation of a drug molecule with a 3D biological molecule like a monoclonal antibody. The isolation and characterization of the siosome conjugates is not as complex as with biological molecules. Scale up GMP production of the siosomes conjugate is easier because it is considered as pharmaceutical drug product. The siosomes conjugates with pharmaceuticals as drug molecules are much more stable than conjugates with biological materials due to risk of e.g. aggregation, degradation, etc. Development and production times for siosomes conjugates are shorter and the costs are cheaper. The siosomes conjugates with drugs are multi-target and delivery systems with better targeting than the monoclonal antibody Drug conjugates This is because of the navigating and targeting molecules on the surface of the siosome. In addition the monoclonal antibodies are designed to target only one antigen or receptor. The documentations required for the marketing authorization of the siosomes conjugates as pharmaceuticals is less comprehensive than for the monoclonal antibody drug conjugates. This will shorten the development time and reduce the costs.

Example 5

(106) 1. Preparation of a Siosome Multi-Target and Delivery of Sugar Organosilicon Compound (Sugar Silane) with Captopril ((S)-1-(3-Mercapto-2-Methyl-1-Oxoropyl)-L-Proline Als ACE2 Inhibitor (Siosomes No. 2—Module 1).

(107) ##STR00018##

(108) Captopril Encapsulated in Siosomes Prepared from Sugar Silane SIL17

(109) A dispersion of 10 μmol of Didodecylsilyl-di(2,3,4,6-O-tetraacetyl-β-D-glucopyranosid) as the representative of amino sugar organosilicon compound, 0.01M Tris/HCl, pH 7.4 and an aqueous solution of Captopril (10 μmol, volume 3 ml) was prepared by mixing with a high pressure homogeniser. The homogenisation time may vary. The mixture was then incubated at 37° C. for 60-90 minutes and sterile filtrated and lyophilised at the temperature of −70° C. The lyophilised-amino sugar organosilicon complex was reconstituted by hydration in sterile (deionized) water and the carrier system solution was used for in-vitro investigations.

(110) After reconstitution by rehydration, the said carrier system surprisingly retains in solution more than 95% of the captopril. The yield of the preparation process resulted from 3 independent experiments, and was 80-90% of the starting concentrations. Stability tests performed at 0, 4, 8, 12 and 48 hours at 4° C., 25° C. and 37° C. have shown that the carrier system complex of organosilicon captopril is surprisingly stable and retains its activity.

(111) The preparation of further siosomes from the modules 1, 2, 3 will be adjusted according to the customized molecular structure. And the residues R1, R2, R3, R4 and the molecules on the surface of the siosomes and/or encapsulated, entrapped and/or conjugated

(112) 2. Specific Multi-Targeting of the GI (Gastro Intestinal Tract)—(FIG. 6)

(113) The oral route is attractive for drug administration because it is associated with patient acceptability, less stringent production conditions, and lower costs. Siosomes as multi-target and delivery system have many suitable properties for increasing the interaction between drugs and the mucosae. and targeting the receptors on the epithelial cells.

(114) SARS Co-2 v and other viruses target and infect the GIT and the siosomes as the multi-target and delivery system will be able to block the ACE2 receptors in the host cells. In addition the inhibition of the virus via the blocking and inhibition of the spike protein (s) and the virion as explained in the functions of the siosomes of module 1, 2 and 3 according to the invention.

(115) Furthermore, according to the invention the multi-target and delivery system could be administered orally, which will be as well of great advantage for the prophylaxis, attenuation, prevention and/or treatment of the infections caused by SARS Co-2 V.

(116) The following specific GI compounds will be directly linked, encapsulated or entrapped in the siosomes: N-Acetyl Cysteine Glutamine Fumarate Glycolic Acid Sodium Glycocholate Sodium deoxy cholate Sodium caprate Lectins Chitosan Poly lactic
3. Specific Targeting the Brain-BBB Using the Siosomes as Multi-Target and Delivery System and CNS Navigators

(117) SARS Co-2 v and other viruses target and infect the CNS. Therefore it is an object of the invention to target the CNS.

(118) Development of therapeutics for brain disorders is one of the more difficult challenges to be overcome by the scientific community due to the inability of most molecules to cross the blood-brain barrier (BBB). Antibody-conjugated nanoparticles are drug carriers that can be used to target encapsulated drugs to the brain endothelial cells and have proven to be very promising. They significantly improve the accumulation of the drug in pathological sites and decrease the undesirable side effect of drugs in healthy tissues. But they have serious problems concerning the very comprehensive development and production and stability of the monoclonal antibody complex.

(119) The siosomes as multi-target and delivery system are pharmaceutical carriers and can provide very specific and stable targeting systems. A number of very specific navigators could be incorporated on the surface of the siosomes and be a part of the therapy strategy of the SARS Co-2 V.

(120) The following neurotransmitters and compounds could be incorporated in the siosomes and as well encapsulated and/or entrapped in the siosomes:

(121) ##STR00019##
Major neurotransmitters are listed below: Amino acids: glutamate, aspartate, D-serine, γ-aminobutyric acid (GABA), glycine Gasotransmitters: nitric oxide (NO), carbon monoxide (CO), hydrogen sulfide (H.sub.2S) Monoamines: dopamine (DA), norepinephrine (noradrenaline; NE, NA), epinephrine (adrenaline), histamine, serotonin (SER, 5-HT) Trace amines: phenethylamine, N-methylphenethylamine, tyramine, 3-iodothyronamine, octopamine, tryptamine, etc. Peptides: oxytocin, somatostatin, substance P, cocaine and amphetamine regulated transcript, opioid peptides Purines: adenosine triphosphate (ATP), adenosine Catecholamines: dopamine, norepinephrine (noradrenaline), epinephrine (adrenaline) Others: acetylcholine (ACh), anandamide, etc

(122) The preparation of the multi-target and delivery systems will be performed according to the procedures of this invention.

Example 6

(123) Preparation of the multi-target and delivery siosomes for the attenuation, prevention and/or treatment of the infections caused by the SARS Co 2-V Virur (FIGS. 14,15).

(124) Examples for the structures of the sugar silanes and the siosomes from the Modules 1, 2, 3.1 and 3.2 are:

(125) 1. As an Example Silane/Siosome No. 2 Module 1 with Encapsulated Captopril (Table 1)

(126) ##STR00020##
2. As an Example Silane/Siosome No. 5 Module 2 with Encapsulated Dipeptide (Table 2)

(127) ##STR00021##
3. As an Example Silane/Siosome No. 3 Module 3.1 with Encapsulated m RNA (Table 3.1)

(128) ##STR00022##
4. As an Example Silane/Siosome No. 1 Module 3.2 with Encapsulated Camostat Mesylate with Cationic Siosomes. Cam=Camostat—(Table 3.2)

(129) ##STR00023##
1. Preparation of any of the multi-targeting and delivery system (siosome) according to any one of the preceding claims, characterized in that the multi-targeting and delivery system is obtainable by: a) mixing one or more organosilicon, sugar organosilicon, amino-sugar organosilicon and/or the vesicles formed from them, with one or more of the active substances at selected pH, salt concentration and temperature; b) homogenisation, sonication and/or extrusion of the mixture, followed by c) separation of the free active substance, d) sterile filtration of the mixture, e) lyophilisation, f) reconstitution to form a siosome of the multi-target and delivery system.
2. The selected (customized) siosomes from module 1, 2, 3 with the defined functions will be prepared above according to the invention separately as lyophilized powder.
3. The lyophilized powder composition comprising siosomes 1, 2, 3 will be mixed at a ratio of X:Y:Z.
4. The mixed lyophilized powder will be used for the preparation of the following formulations: Oral, rectal, vaginal, topical, nasal, intradermal, or parenteral administration (FIG. 15).