Antimicrobial fusion compounds and uses thereof

Abstract

A fusion protein comprising at least one Type 1 Ribosome Inactivating Protein, polypeptide B; and at least one polypeptide A capable of viral entry inhibition; and/or at least one Cationic AntiMicrobial Peptide, polypeptide C.

1 . A fusion protein comprising at least one Type 1 Ribosome Inactivating Protein (RIP) or fragment thereof, polypeptide B; and
 (i) at least one polypeptide A capable of viral entry inhibition; and/or   (ii) at least one Cationic AntiMicrobial Peptide (CAP) or fragment thereof, polypeptide C,   
       wherein the fusion protein is recombinant and comprises at least one linker peptide between each of the polypeptides A, B and C. 
     
     
         2 . The fusion protein according to  claim 1 , wherein the polypeptide A is theta defensin, an analogue, or a fragment thereof. 
     
     
         3 . The fusion protein according to  claim 1 , wherein the fusion protein comprises the structure A-B-C, A-C-B, C-A-B, C-B-A, B-A-C, B-C-A, A-B-C-C, A-B, B-A, B-C, C-B, C-B-C, or C-C-B-C-C. 
     
     
         4 . The fusion protein according to  claim 1 , wherein the fusion protein comprises polypeptides A, B and C. 
     
     
         5 . The fusion protein according to  claim 1 , wherein polypeptides A, B and C are directly linked to one another. 
     
     
         6 . (canceled) 
     
     
         7 . The fusion protein according to  claim 1 , wherein the linker peptide has SEQ ID NO: 11. 
     
     
         8 . The fusion protein according to  claim 1 , wherein polypeptide A is selected from the group consisting of Rhesus minidefensin (RTD-1), RTD-2, RTD-3, Retrocyclin-1, Retrocyclin-2, Retrocyclin-3, synthetic retrocyclin congener RC100, RC101, RC102, RC103, RC104, RC105, RC106, RC107, RC108, RC110, RC111, RC112, RC113 and RC114. 
     
     
         9 . The fusion protein according to  claim 1 , wherein the Type 1 RIP (polypeptide B) is selected from the group consisting of Ebulitins, Nigritins, Amarandins,  Amaranthus antiviral/ RIP, Amaranthin,  Atriplex patens  RIP,  Beta vulgaris  RIP, β-vulgin,  Celosia cristata  RIP,  Chenopodium album  RIP, CAP30B,  Spinacea oleracea  RIP, Quinqueginsin, Asparins, Agrostin, Dianthins, DAPs,  Dianthus chinensis',  Lychnin, Petroglaucin, Petrograndin,  Saponaria ocymoides  RIP,  Vacuolas saporin,  Saporins,  Vaccaria hispanica  RIP, Benincasins, Hispin, Byrodins, Colocins,  Cucumis figarei  RIP, Melonin,  C. moschata  RIP, Cucurmosin, Moschatins, Pepocin, Gynostemmin,  Gynostemma pentaphyllum  RIP, Gypsophilin, Lagenin, Luffaculin, Luffangulin, Luffin, MORs, Momordin II, Momorcharins, Momorcochin, Momorcochin-S, Sechiumin, Momorgrosvin, Trichoanguin, Kirilowin, α-trichosanthin, TAP-29, Trichokirin, Trichomislin, Trichosanthin, Karasurin, Trichomaglin, Trichobakin, Crotin, Euserratin Antiviral Protein GAP-31, Gelonin,  Hura crepitans  RIP, Curcin,  Jathropa curcas  RIP, Mapalmin, Manutins, α-pisavin, Charibdin,  Hyacinthus orientalis  RIP, Musarmin,  Iris hollandica  RIP,  Cleroendrum aculeatum  RIP, CIPs), Crip-31, Bouganin,  Bougainvilla spectbilis  RIP,  Bougainvillea×buttiana  Antiviral protein 1 (BBAP1), Malic enzymes, MAP-S, pokeweed antiviral proteins (PAP), PD-SI, DP-S2, Dodecandrin, PIP, PIP2,  Phytolacca octandra  anti-viral proteins,  Hordeum vulgare  RIP's,  Hordeum vulgare  sub sp.  Vulgare  Translational inhibitor II,  Secale cereale  RIP, Tritin,  Zea diploperemis  RIPs,  Malus×domestica  RIP, Momordica Anti-HIV Protein,  Gelonium multiflorum, Mirabilis expansa  1, phage MU1, betavulgin (Bvg), curcin 2, saporin 6, Maize RIP (B-32), Tobacco RIP (TRIP), Beetins,  Mirabilis  antiviral protein (MAP), Trichosanthin (TCS), luffins, Momorcharins, Ocymoidin, Bryodin, Pepopsin, β-trichosanthin, Camphorin, YLP, Insularin, Barley RIP, Tritins, Lamjarin, and  Volvariella volvacea  RIP. 
     
     
         10 . The fusion protein according to  claim 1 , wherein the CAP (polypeptide C) is selected from the group consisting of Cyclotides, Siamycins, NP-06, Gramicidin A, Circulins, Kalatas, Ginkbilobin, Alpha-Basrubin, Lunatusin, Sesquin, Tricyclon A, Cycloviolacins, Polyphemusins, hfl-B5, Protegrins (Pig Cathelicidin), Rat Defensins, Human β-defensins, Temporins, Caerins, Ranatuerins, Reptile Defensin, Piscidins, Lactoferricin B, Rabbit Neutrophils, Rabbit α-Defensin, Retrocyclins, Human α-Defensins, Human β-defensin III (HBD3), Rhesus minidefensin (RTD-1,θ-defensin), rhesus θ-defensins, Human neutrophil peptides, Cecropin As, Melittin, EP5-1, Magainin 2s, hybrid (CE-MA), hepcidin TH1-5, Epinecidin-1, Indolicidin, Cathelicidin-4, LL-37 Cathelicidin, Dermaseptins, Maximins, Brevinins, Ranatuerins, Esculentins, Maculatin 1.3, Maximin H5 and Piscidins, Mundticin KS Enterocin CRL-35, Lunatusin, FK-13 (GI-20 is a derivative), Tachyplesins, Alpha-MSH, Antiviral protein Y3, Palustrin-3AR, Ponericin L2, Spinigerin, Melectin, Clavanin B, Cow cathelicidins, Guinea pig cathelicidin CAP11, Sakacin 5×, Plectasin, Fungal Defensin, GLK-19, lactoferrin (Lf) peptide 2, Alloferon 1, Uperin 3.6, Dahlein 5.6, Ascaphin-8, Human Histatin 5, Guineapig neutrophils, Mytilins, EP5-1, Hexapeptide (synthetic) Corticostatin IV Rabbit Neutrophil 2, Aureins, Latarcin, Plectasin, Cycloviolins, Vary Peptide E, Palicourein, VHL-1, and Buforins. 
     
     
         11 . The fusion protein according to  claim 1 , wherein the Type 1 RIP is MAP30, the CAP a Dermaseptin and the polypeptide A a Retrocyclin. 
     
     
         12 . The fusion protein according to  claim 11 , wherein the Dermaseptin is Dermaseptin 1 and the Retrocyclin is Retrocyclin 101. 
     
     
         13 . The fusion protein according to  claim 1 , comprising the amino acid sequence SEQ ID NO:1. 
     
     
         14 . The fusion protein according to  claim 1 , suitable for oral administration. 
     
     
         15 . An isolated nucleic acid molecule capable of expressing the fusion protein according to  claim 1 . 
     
     
         16 - 17 . (canceled) 
     
     
         18 . A process of producing a fusion protein according to any one of  claim 1 , comprising culturing a host cell comprising an isolated nucleic acid molecule capable of expressing the fusion protein according to  claim 1  under conditions such that the fusion protein is expressed. 
     
     
         19 . A pharmaceutical composition comprising a fusion protein according to  claim 1 . 
     
     
         20 . The pharmaceutical composition according to  claim 19 , wherein the composition is suitable for oral administration. 
     
     
         21 . The pharmaceutical composition according to  claim 19 , further comprising at least one pharmaceutically acceptable carrier, excipient, diluent and/or detergent. 
     
     
         22 - 23 . (canceled) 
     
     
         24 . A method of treating and/or preventing a microbial infection in a vertebrate or invertebrate in need thereof, comprising administering to the vertebrate or invertebrate an effective amount of the fusion protein according to  claim 1 . 
     
     
         25 - 31 . (canceled)