WO2021133928A1

WO2021133928A1 - Antisense oligonucleotides for the treatment of pseudomonas and acinetobacter infections

Info

Publication number: WO2021133928A1
Application number: PCT/US2020/066875
Authority: WO
Inventors: Nrusingh Prasad MOHAPATRA; Denis K. GUENETTE; Kristin Maria KNIGHT; Joseph SALAMOUN
Original assignee: TECHULON Inc
Current assignee: TECHULON Inc
Priority date: 2019-12-27
Filing date: 2020-12-23
Publication date: 2021-07-01
Anticipated expiration: 2022-06-27

Abstract

Disclosed are compounds useful for the treatment of Pseudomonas and Acinetobacter infection. Also disclosed is a pharmaceutical composition comprising the compound and a pharmaceutically acceptable carrier. Also disclosed are methods for the treatment of Pseudomonas and Acinetobacter infections.

Description

ANTISENSE OLIGONUCLEOTIDES FOR THE TREATMENT OF PSEUDOMONAS AND ACINETOBACTER INFECTIONS BRIEF SUMMARY OF THE INVENTION [0001] Provided are PPNA compounds as set forth in Table 1. Table 1: Compounds of the invention

wherein Lx is a linker, X is -NH-(CH2)5C(O)-, R is Arg, hR is homoArg, and x is 1, 2 or 3, or a pharmaceutically acceptable salt thereof. [0002] In some embodiments, L_x comprises a polyethylene glycol (PEG) linker of 2-10 repeating units. In other embodiments, Lx comprises a PEG linker of 6 repeating units. In other embodiments, Lx is -NH-PEG-CH2CH2C(O)-, wherein PEG is polyethylene glycol of 2-10 repeating units; -NH-PEG-CH₂C(O)-, wherein PEG is of 2-10 repeating units; or -NH-PEG-CH2CH2NHC(O)CH2CH2C(O)-, wherein PEG is of 2-10 repeating units. In other embodiments, the polynucleotide of the compound comprises a modified backbone. [0003] In some embodiments, the compound is depicted in Fig 2, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 3)-, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 4)-, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 5, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 6, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 7, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 8, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 9, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 10, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 11, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 12, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 13, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 14, or a pharmaceutically acceptable salt thereof. In other embodiments, the compound is depicted in Fig 15, or a pharmaceutically acceptable salt thereof. [0004] Also provided is a pharmaceutical composition comprising the compound described herein and a pharmaceutically acceptable carrier. [0005] Also provided is a method of treating a bacterial infection, comprising administering to an animal in need thereof an effective amount of a compound or pharmaceutical composition as described herein. [0006] In some embodiments, the bacteria is Pseudomonas and/or Acinetobacter. In other embodiments, the bacteria is Pseudomonas aeruginosa. In other embodiments, the bacteria is Acinetobacter baumannii. [0007] In some embodiments, the animal has hospital/ventilator associated pneumonia. In other embodiments, the animal has sepsis. In other embodiments, the animal has a urinary tract infection. In other embodiments, the animal has soft tissue-associated infection. In other embodiments, the animal has hardware prosthetic-associated infection. [0008] In some embodiments, the treatment route comprises intravenous, intraperitoneal or oral administration. [0009] In some embodiments, the animal is a human. BRIEF DESCRIPTION OF THE FIGURES [0010] Fig. 1 is a table listing of the compounds disclosed in the invention, wherein Lx is a linker, X is -NH-(CH2)5C(O)-, R is Arg, hR is homoArg, and x is 1, 2 or 3, or a pharmaceutically acceptable salt thereof. [0011] Fig. 2 is a depiction of the structure of PPNA 40-PEG6. [0012] Fig. 3 is a depiction of the structure of PPNA 39-PEG6. [0013] Fig. 4 is a depiction of the structure of PPNA 38-PEG6. [0014] Fig. 5 is a depiction of the structure of PPNA 36-PEG6. [0015] Fig. 6 is a depiction of the structure of PPNA 23X-PEG6-CPP2. [0016] Fig. 7 is a depiction of the structure of rpIR-PEG6-CPP2. [0017] Fig. 8 is a depiction of the structure of rpIV-PEG6-CPP2. [0018] Fig. 9 is a depiction of the structure of pheA-PEG6-CPP3. [0019] Fig. 10 is a depiction of the structure of rpIR-PEG6-CPP3. [0020] Fig. 11 is a depiction of the structure of pheA-PEG6-CPP5. [0021] Fig. 12 is a depiction of the structure of PPNA 40-PEG6-Homoarginine. [0022] Fig. 13 is a depiction of the structure of PPNA 39-PEG6-Homoarginine. [0023] Fig. 14 is a depiction of the structure of PPNA 38-PEG6-Homoarginine. [0024] Fig. 15 is a depiction of the structure of PPNA 36-PEG6-Homoarginine. [0025] Fig. 16 is a depiction of the structure of PPNA40-PEG3. [0026] Fig. 17 is a bar chart showing the efficacy of different amounts of PPNA 23X- PEG6-CCPS following intravenous, intraperitoneal, and subcutaneous dosing in the neutropenic mice thigh infection model against P. aeruginosa. [0027] Fig. 18 is a bar chart showing the antimicrobial efficacy different amounts of PPNA 25X-PEG6-CCPS following intravenous, intraperitoneal, and subcutaneous dosing in the neutropenic mice thigh infection model against P. aeruginosa. DETAILED DESCRIPTION OF THE INVENTION [0028] The present invention may be understood by reference to the following detailed description of the embodiments of the invention herein. The terminology used herein is for the purpose of describing embodiments of the invention and is not intended to be limiting. [0029] Specific aspects of the invention include a method of treating a bacterial infection and/or inhibiting growth of Pseudomonas and/or Acinetobacter. In some embodiments, the bacterial infection is caused by Pseudomonas aeruginosa and/or Acinetobacter baumannii. [0030] The compounds of the invention comprise an antisense molecule that inhibits the growth of Pseudomonas and/or Acinetobacter, a linker, and a cell penetration peptide (CPP). The cell penetration peptide may have one or more functions to facilitate cell targeting and/or membrane permeation of a bacteria in a host. The cell penetration peptide provides for membrane disruption of bacteria with specificity and reduces toxicity. [0031] Bulk synthesis can be carried out by contract manufacturers, such as Bachem Inc. (Torrence, CA) or AmbioPharm, Inc. (North Augusta, SC) or CPC Scientific Inc. (San Jose, CA) using standard methodologies including solid-scaffold protection/deprotection synthesis via high fidelity synthesizers. [0032] In one embodiment of the invention, the compound is part of a composition comprising a buffer. Suitable buffers in the composition of the invention provide a basic pH when dissolved or dispersed in water. In some embodiments, the buffer has a pKa of greater than about 7. See, for example, "Handbook of Pharmaceutical Excipients," 5^th ed., Rowe et al. (eds.) (2006); and SIGMA Life Sciences, "Products for Life Science Research," Product Catalog (2008-2009). The composition may comprise one or more buffers. Such buffers include—but are not limited to—phosphate buffers, bicarbonate buffers, ethanolamine buffers, borate buffers, imidazole buffers, tris buffers, and zwitterionic buffers (e.g., HEPES, BES, PIPES, Tricine, and other so-called "Good's Buffers"). See, for example, Good et al., "Hydrogen Ion Buffers for Biological Research," Biochemistry, 5(2):467–477 (1966). In one particular embodiment, the buffer is a bicarbonate, such as sodium bicarbonate or carbonate. [0033] In one embodiment of the invention, the buffer has a pKa between about 6 and about 14, between about 7 and about 13, and between about 7 and about 12. In another embodiment, the buffer has a pKa between about 7 and about 8. [0034] In another embodiment of the invention, the compound is combined with a delivery polymer. The polymer-based nanoparticle drug delivery platform is adaptable to a diverse set of polynucleotide therapeutic modalities. In one aspect of the invention, the delivery polymer is cationic. In another aspect of the invention, the delivery polymer comprises phosphonium ions and/or ammonium ions. In another example of the invention, the compound is combined with a delivery polymer, and the composition forms nanoparticles in solution. In a further embodiment, nanoparticle polyplexes are stable in serum and have a size in the range of about 30 nm – 5000 nm in diameter. In one embodiment, the particles are less than about 300 nm in diameter. For example, the nanoparticles are less than about 150 nm in diameter. [0035] In one embodiment, the delivery vehicle comprises a cationic block copolymer comprising phosphonium or ammonium ionic groups as described in PCT/US12/42974. In one embodiment, the polymer is diblock-Poly[(ethylene glycol)₉ methyl ethyl methacrylate][stearylphosphonium]. In another embodiment of the invention, the delivery polymer comprises glycoamidoamines as described in Tranter et al. Amer Soc Gene Cell Ther, Dec 2011; polyhydroxylamidoamines, dendritic macromolecules, carbohydrate- containing polyesters, as described in US20090105115; and US20090124534. In other embodiments of the invention, the nucleic acid delivery vehicle comprises a cationic polypeptide or cationic lipid. An example of a cationic polypeptide is polylysine. See U.S. Pat. 5,521,291. [0036] In one embodiment, the compound is part of a composition comprising delivery or carrier polymers. In another embodiment, the compound is part of nanoparticle polyplexes capable of transporting molecules with stability in serum. The polyplex compositions comprise a synthetic delivery polymer (carrier polymer) and biologically active compound associated with one another in the form of particles having an average diameter of less than about 500 nm, such as about 300 nm, or about 200 nm, preferably less than about 150 nm, such as less than about 100 nm. The invention encompasses particles in the range of about 40 nm – 500 nm in diameter. [0037] In one embodiment, the delivery or carrier polymer comprises a cationic block copolymer containing phosphonium or ammonium ionic groups as described in PCT/US12/42974. In another embodiment of the invention, the delivery or carrier polymer comprises glycoamidoamines as described in Tranter et al. Amer Soc Gene Cell Ther, Dec 2011; polyhydroxylamidoamines, dendritic macromolecules, carbohydrate- containing polyesters, as described in US20090105115; and US20090124534. The polyglycoamidoamine (PGAA) polymer system, which is a proprietary, localized and biodegradable nanoparticle system, represents another delivery or carrier polymer. Poly(galactaramidoamine) is an efficient cationic polymeric vehicle with low cytotoxicity (Wongrakpanich et al. Pharmaceutical Development and Technology, January 12, 2012). The nanoparticle delivery system disclosed in Hemp et al. Biomacromolecules, 2012 13:2439-45 represents another delivery or carrier polymer useful in the present invention. [0038] In other embodiments of the invention, the delivery or carrier polymer comprises a cationic polypeptide or cationic lipid. Polymers, such as poly-L-lysine (PLL), polyethyleneimine (PEI), chitosan, and their derivatives are also encompassed by the invention. Nucleic acid delivery using these compounds relies on complexation driven by electrostatic interactions between the gene and the polycationic delivery agent. Polymer- DNA complexes condense into particles on the order of 60 nm – 120 nm in diameter. Polymers such as linear PEI and PLL have high transfection rates in a variety of cells. [0039] In vivo nucleic acid delivery has size constraints requiring a sufficiently small polyplex to enable long circulation times and cellular uptake. In addition, polyplexes must resist salt- and serum-induced aggregation. Serum stability is generally associated with a particle size of about sub-150 nm hydrodynamic radius or below maintainable for 24 h. The nanoparticles of the invention, which comprise nucleic acid therapeutic and delivery polymer, have the hydrodynamic radius and material properties for serum stability. In particular, the delivery polymer, when combined with the nucleic acid, protects the therapeutic cargo under physiological conditions. The delivery polymers are designed to have characteristics of spontaneous self-assembly into nanoparticles when combined with polynucleotides in solution. [0040] The invention also contemplates other delivery polymers that form serum-stable nanoparticles. The invention is not limited to the type of delivery polymer and may be adaptable to nucleic acid characteristics, such as length, composition, charge, and presence of coupled peptide. The delivery polymer may also be adaptable for material properties of the resultant nanoparticle, such as hydrodynamic radius, stability in the host bloodstream, toxicity to the host, and ability to release cargo inside a host cell. [0041] In one embodiment, the compound is administered in the form of a salt. The salt may be any pharmaceutically acceptable salt comprising an acid or base addition salt. Examples of pharmaceutically acceptable salts with acids include those formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, and the like. Also included are salts that are formed with organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. and include, for example, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p- toluenesulfonic acid, salicylic acid, and the like. Exemplary salts thus include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, trifluoroacetates, propionates, caprylates, isobutyrates, oxalates, malonates, succinate suberates, sebacates, fumarates, maleates, mandelates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, malates, tartrates, methanesulfonates, and the like. Also contemplated are salts of amino acids, such as arginates, gluconates, and galacturonates (see, for example, Berge S. M. et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 66:1-19 (1997). Acid addition salts of basic molecules may be prepared by contacting the free base forms with a sufficient amount of the desired acid to produce the salt according to methods and techniques with which a skilled artisan is familiar. [0042] Pharmaceutically acceptable base addition salts are formed by addition of an inorganic base or an organic base to the free acid. Pharmaceutically acceptable base addition salts may be formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2- diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, N,N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline, N-methylglucamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like. [0043] In one embodiment, the compound is administered as part of a pharmaceutical composition comprising a pharmaceutically acceptable diluent, excipient or carrier. Suitable diluents, excipients and carriers are well known in the art and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A.R. Gernnaro Ed., 1985). The pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the ultimate dosage form must be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, saline, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin. [0044] Sterile injectable solutions are prepared by incorporating the compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions. [0045] In one embodiment, the composition comprising the compound is in contact with a fabric. The fabric may comprise natural fibers, synthetic fibers, or both. Examples of textile fabrics include, but are not limited to, nylon, cotton, nylon-cotton blends, wool, silk, linen, polyester, rayon, and worsted. In one particular embodiment of the invention, the fabric is cotton. In another embodiment, the fabric is nylon. In another embodiment, the fabric is a nylon-cotton blend. The ratio of nylon to cotton in the nylon-cotton blend fabric can be between about 1:99 and about 99:1, between about 10:90 and about 90:10, between about 20:80 and about 80:20, between about 30:70 and about 70:30, between about 40:60 and about 60:40, and between about 45:55 and about 55:45. In a preferred embodiment, the fabric is a 50:50 nylon-cotton blend. [0046] In another embodiment of the invention, the fabric has a high tensile strength-to- weight ratio. In one embodiment, the fabric with a high tensile-to-weight ratio is a fabric comprising aramid fibers. In a particular embodiment, the aramid fiber is a para-aramid fiber (e.g., the para-aramid fiber commercially known as KEVLAR). In another particular embodiment, the aramid fiber is a meta-aramid fiber (e.g., the meta-aramid fiber commercially known as NOMEX). [0047] In certain embodiments, the antimicrobial fabric is capable of treating a Pseudomonas and/or Acinetobacter infection or inhibiting growth of Pseudomonas and/or Acinetobacter after the fabric has been washed. In some embodiments, the bacterium is Pseudomonas aeruginosa. In some other embodiments, the bacterium is Acinetobacter baumannii. In some embodiments, the antimicrobial fabric is capable of treating a Pseudomonas and/or Acinetobacter infection or inhibiting growth of Pseudomonas and/or Acinetobacter after between about 10 and about 60 wash cycles, between about 20 and about 50 wash cycles, between about 20 and about 40 wash cycles, between about 20 and about 30 wash cycles, and between about 20 and about 25 wash cycles. In another embodiment, the duration of a wash cycle is between about 10 minutes and about 90 minutes, between about 10 minutes and about 75 minutes, between about 10 minutes and about 60 minutes, between about 10 minutes and about 45 minutes, between about 10 minutes and about 30 minutes, and between about 10 minutes and about 15 minutes. In another embodiment, the water temperature in the wash cycles is between about 16°C and about 60°C, between about 27°C and about 49°C, or between about 37° and about 44°C. In one particular embodiment, the antimicrobial fabric is capable of treating a Pseudomonas and/or Acinetobacter infection or inhibiting growth of Pseudomonas and/or Acinetobacter following Laundry Test Method AATCC 147 from American Association of Textile Chemists and Colorists (AATCC). [0048] In another embodiment, provided is a composition comprising the compound. The composition may be in the form of solution that can be applied to a fabric, e.g., by rinsing, dipping, or spraying. The fabric can be an antimicrobial fabric or a non- antimicrobial fabric. In one embodiment, application of the solution to the fabric provides a fabric that is capable of treating a Pseudomonas and/or Acinetobacter infection or inhibiting growth of Pseudomonas and/or Acinetobacter. In other embodiments, application of the solution to the fabric increases the fabric's capability of treating a Pseudomonas and/or Acinetobacter infection or inhibiting growth of Pseudomonas and/or Acinetobacter. In a particular embodiment, application of the solution to an antimicrobial fabric with low antimicrobial activity increases the antimicrobial activity of the fabric. [0049] In other embodiments of the invention, provide is a wound healing dressing comprising the compound. In one embodiment, the wound healing dressing is an adhesive dressing. In another embodiment, the wound healing dressing is a non-adhesive dressing. In one embodiment, the dressing comprises a foam, gel, or cream. In another embodiment, the dressing comprises a fiber based material (e.g., gauzes or waddings). In one embodiment, the fiber-based material is cotton. In another embodiment, the fiber- based material is rayon. In another embodiment, the fiber-based material is a gel-forming fiber, such as a carboxymethylated cellulosic material. In another embodiment, the fiber- based material is a synthetic polymer. In another embodiment, the wound healing dressing is THERAGAUZE (Soluble Systems, LLC, Newport News, VA). [0050] The invention also provides a method of treating Pseudomonas and/or Acinetobacter infection and a method of inhibiting growth of Pseudomonas and/or Acinetobacter. In one embodiment, the animal undergoing treatment for Pseudomonas and/or Acinetobacter infection exhibits one or more symptoms of Pseudomonas and/or Acinetobacter infection including fever and chills, body aches, light-headedness, rapid pulse and breathing, nausea and vomiting, diarrhea, infections of the lungs, blood, brain, urinary track, and wound infections, fever, confusion, neck stiffness, sensitivity to light, bloodstream infection, fever, chills, rash, light-headedness, altered mental state as well as puss production in the infected area, acne, boils, abscesses, carbuncles, stys, cellulitis, diarrhea, botulism, and gas gangrene. The animal may also exhibit signs of sepsis or pneumonia. [0051] In one embodiment, the compound is administered by intravenous injection. In another embodiment, the compound is administered by intramuscular injection. In another embodiment, the compound is administered by peritoneal injection. In another embodiment, the compound is administered topically, e.g. to a tissue suspected to be infected by Pseudomonas and/or Acinetobacter. In another embodiment, the compound is administered orally. When administered orally, the compound may be formulated as part of a pharmaceutical composition coated with an enteric coating that will protect the compound from the acid environment of the stomach and release the compound in the upper gastrointestinal tract. In another embodiment, the compound may be formulated as part of a sustained release formulation that will release the compound on a substantially continuous basis over a period of time. [0052] Animals that may be treated with the compound according to the invention include any animal that may benefit from treatment with the compound. Such animals include mammals such as humans, dogs, cats, cattle, horses, pigs, sheep, goats and the like. [0053] The compound is administered in an amount that is effective for the treatment of Pseudomonas and/or Acinetobacter infection or inhibition of the growth of Pseudomonas and/or Acinetobacter. The amount may vary widely depending on the mode of administration, the species of Pseudomonas and/or Acinetobacter, the age of the animal, the weight of the animal, and the surface area of the animal. The amount of compound, salt and/or complex thereof may range anywhere from 1 pmol/kg to 1 mmol/kg. In another embodiment, the amount may range from 1 nmol/kg to 10 mmol/kg. When administered topically, the amount of compound, salt and/or complex thereof may range anywhere from 1 to 99 weight percent. In another embodiment, the amount of compound, salt and/or complex thereof may range anywhere from 1 to 10 weight percent. [0054] The invention also provides compounds comprising a linker. In one embodiment, the compound is an antisense molecule that inhibits the growth of a bacterium comprising a polynucleotide sequence that is antisense to the coding region of a bacterial protein and hybridizes to the coding region under physiological conditions; bound to a linker that has a formula (Y')n, where each Y' is independently glycine, cysteine, 8-amino-3,6- dioxaoctanoic acid (AEEA), or 5-amino-3-oxapentanoic acid (AEA), and n is an integer from 1 to 10; and bound to a cell penetrating molecule (CPP). In some aspects, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In another embodiment, linker is a bond. In another embodiment, the linker is a polyethylene glycol (PEG) of 2-10 repeating units. In another embodiment, the PEG has 2, 3, 4, 5, 6, 7, 8, 9 or 10 repeating units. [0055] In some aspects, the PCC has the formula: (ABC)p-D, wherein A is a cationic amino acid which is Lysine or Arginine; B and C are hydrophobic amino acids which may be the same or different and are selected from the group consisting of Valine, Leucine, Isoleucine, Tyrosine, Phenylalanine, and Tryptophan; p is an integer with a minimal value of 2; and D is a cationic amino acid or is absent. In one embodiment, A is Lysine, B is Phenylalanine, C is Phenylalanine, D is Lysine, and p is 3. In another embodiment, p is 2-10. In another embodiment, p is 2, 3, 4, 5, 6, 7, 8, 9 or 10. [0056] In other aspects, the PCC has the formula: B-X1-(R-X2-R)4, where B is beta- alanine or is absent, X₁ is 6-amino-hexanoic acid or is absent, X₂ is 6-amino-hexanoic acid, and R is arginine or homo-arginine. In some embodiments, R is arginine, selected from the group consisting of L-arginine and D-arginine. [0057] In some aspects, the PCC is a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-15 (See Table 2). Table 2: Cell Penetrating Peptides

[0058] In Table 2, X is -NH-(CH₂)₅C(O)- and R is L-Arg or homoArg. [0059] In some embodiments, the polynucleotide of the compound comprises a modified backbone. In a particular embodiment, the modified backbone is a PNA backbone. [0060] Modified nucleic acids are non-natural polymers that hybridize to natural DNA and RNA with sequence specificity according to Watson-Crick base paring rules. Examples of modified nucleic acids are phosphorothioate-oligodeoxynucleotides (PS- ODNs), locked nucleic acids (LNAs), 2'-O-methyloligoribonucleotides (2'O-Mes), phosphorodiamidate morpholino oligonucleotides (PMOs), and peptide nucleic acids (PNAs). Modified nucleic acids have modified backbones and are generally more resistant to degradation than natural nucleic acids. The invention includes any type of synthetically-modified DNA or RNA that hybridizes to natural DNA and RNA. See, e.g., U.S. Pat. Nos. 5,116,195, 5,539,082, 5,527,675, 5,623,049, 5,714,331, 5,736,336, 5,773,571, 5,786,461, 5,811,232, 5,837,459, 5,874,564, 5,891,625, 5,972,610, 5,986,053, 6,107,470, 6,174,870, 7,098,192, 7,696,345, 8,124,745, 8,354,093, 8,357,664, Wagner et al., Nucl. Acid Res. 19:5965-71 (1991); and Koshkin et al., Tetrahedron 54:3607-30 (1998). [0061] Antisense molecules of the invention may also be composed of non-natural polymers that hybridize to natural nucleic acids. Atypical nucleoside bases may also be employed, such as methylated bases, phosphorylated bases, inosine, thiouridine, pseudouridine, dihydrouridine, queuosine, and wyosine, among others. Examples of such antisense polymers comprising atypical bases are disclosed in U.S. Pat. Nos. 7,875,733, 7,919,612, 7,939,677, 8,314,229, 8,372,969, and 8,377,898. [0062] The term antisense polynucleotide refers to a nucleic acid molecule that is complementary to at least a portion of a target nucleotide sequence of interest and hybridizes to the target nucleotide sequence under physiological conditions. Antisense molecules specifically hybridize with one or more nucleic acids encoding a preselected target nucleic acid. The terms target nucleic acid and nucleic acid encoding the target encompass DNA encoding the target, RNA (including pre-mRNA and mRNA) transcribed from such DNA, and also cDNA derived from such RNA. The hybridization of an antisense compound with its target nucleic acid interferes with the normal function of the nucleic acid. This modulation of function of a target nucleic acid by compounds which specifically hybridize to it is generally referred to as antisense. The functions of DNA to be interfered with include replication and transcription. The functions of RNA to be interfered with include all vital functions such as, for example, translocation of the RNA to the site of protein translation, translation of protein from the RNA, splicing of the RNA to yield one or more mRNA species, and catalytic activity which may be engaged in or facilitated by the RNA. The overall effect of such interference with target nucleic acid function is modulation of the expression of the target. In the context of the present invention, modulation means either an increase (stimulation) or a decrease (inhibition) in the expression of a gene. In the context of the present invention, inhibition is the form of modulation of gene expression. [0063] Polynucleotides are described as complementary to one another when hybridization occurs in an antiparallel configuration between two single-stranded polynucleotides. [0064] The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity and similarity between two sequences can be accomplished using a mathematical algorithm (see e.g., Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991). In a preferred embodiment, the percent identity between two sequences is determined based on alignments generated with the Clustal W algorithm (Thompson, J. D. et al., 1994, Nucleic acids Res. 22:4673-4680). This algorithm is incorporated into many commercial software packages, in this case the alignX software program in the Vector NTI suite (version 8.0). Default Clustal W parameters were used to generate pairwise alignments from which percent identity values were calculated (gap opening penalty of 10; gap extension penalty of 0.1). The percent identity is defined as the number of identical bases divided by the total number of bases and multiplied by 100. If sequences in the alignment are of different lengths (due to gaps or extensions), the length of the longest sequence will be used in the calculation, representing the value for total length. [0065] Proteins are polymers containing one or more chains of amino acids bonded together by peptide bonds. Proteins typically fold into a three dimensional form, facilitating a biological function. [0066] A polypeptide is a polymer of amino acids bonded together by peptide bonds. The terms protein and polypeptide and peptide are generally used interchangeably, although polypeptides and peptides are generally shorter in length than proteins. [0067] The invention provides compounds which are used for the treatment of infections on an animal caused by Pseudomonas and/or Acinetobacter. In some embodiments, the bacteria are Pseudomonas aeruginosa and/or Acinetobacter baumannii. In one embodiment, the animal has hospital/ventilator associated pneumonia. In another embodiment, the animal has sepsis. In one embodiment, the animal has a urinary tract infection (UTI). In another embodiment, the animal has soft tissue-associated infection. In another embodiment, the animal has hardware prosthetic-associated infection. [0068] The treatment routes of the invention include, but are not limited to, IV, IP, and oral. [0069] The invention provides the specific compounds shown in Figures 1-15. EXAMPLE Example 1 Synthesis of PNA-CPP derivatives [0070] PNA-CPP derivatives of the present disclosure were synthesized by following Merrifield Solid Phase Peptide Synthesis using AAPPTEC automated peptide synthesizers or manually. Each compound was synthesized up to 1.0 micromolar (µM) concentration using Rink-Amide™ or Tentagel XV-RAM™ resin in 50-500 ml reaction vessels. The Rink-amide™ or Tentagel XV-RAM™ resins were deprotected by 20% piperidine in N-Methyl-2-pyrrolidone (NMP). Resins were washed with NMP for 7 times with 2 mins mixing. Five equimolar concentration of Fmoc-amino acids/Fmoc-PNAs were mixed with 4.75 equimolar concentrations of 1-Cyano-2-ethoxy-2- oxoethylidenaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate in presence of NMP for 1 min and added to the deprotected resins with further addition of 0.5M N,N-Diisopropylethylamine (DIEA) and 0.5M of 2,6-lutidine for coupling of Fmoc- amino acids/Fmoc-PNAs. Coupling of Fmoc-amino acids/Fmoc-PNAs was performed for 30 mins with continuous shaking, intermittent argon gas bubbling at 50^oC. The coupling reaction was repeated twice, and resins were washed five times with NMP with 1 mins mixing at 50^oC. The growing amino acids/PNAs on resins were capped with 2.5 M Acetic anhydride for 30 mins followed by 5 times washing of resins with NMP with 2 mins of shaking with no heating. The process of deprotection, coupling, and capping steps repeated till the end of synthesis of compound. After final capping of amino acid/PNA onto growing resins, the final product was deprotected from resins. The crude product was cleaved from resin by 95% trifluoroacetic acid, 2.5% TIS and 2.5 water for 4 hrs at room temperature. The cleavage product was precipitated in 10 volumes of cold ether and the precipitated compound was collected by centrifugation. The ether precipitated compound was air dried for purification. Example 2 Purification of the PNA-CPP derivatives [0071] The air dried crude compounds were solubilized in 0.1% TFA in HPLC water. The compounds were purified in Waters Prep-150 system. Thirty milligrams of compound was loaded in X-Bridge™ C18 columns (10mm X 250mm) with a flow rate of mobile phase 7 ml/min. Mobile phase conditions are as follows in Table 3: Table 3: Mobile Phase Conditions for Purification of PNA-CPP Conjugates

[0072] The purified fractions were lyophilized and converted to acetate salts. The acetate salt of the compounds were converted by passing through the HPLC columns in a 10% acetic acid-water and acetonitrile mobile phase. The purified fractions were lyophilized to collect the acetate salts of the compounds. A small fraction of the compound was used to run in an analytical HPLC column to determine the purity of the compound. The purity of all compounds were >95%. The acetate salt of compounds were screened for the antimicrobial activities against bacterial strains. [0073] Table 4: Provided are compounds as depicted in the following table: Table 4

a-Adenine; g-Guanine; t-Thymine; c-Cytosine; R- L-Arginine; K- L-Lysine; F- L- phenylalanine; A-L-Alanine; I-L-Isoleucine; L-L-Leucine; W-L-Tryptophan; Y-L-Tyrosine; hR- Homo-Arginine; X- 6-amino-hexanoic acid Example 3 Assays to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) PNA-CPP derivatives against P. aeruginosa and A. baumannii strains. [0074] Minimum inhibitory concentration (MIC) analyses were performed as described in Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically.11th ed. CLSI standard M07. Wayne, PA: Clinical and Laboratory Standards Institute; 2018 to detect the MIC and MBC of PNA-CPP derivatives against P. aeruginosa and A. baumannii strains. Colistin was used as control. Different concentration of PNA-CPP derivatives were prepared in 10mM of sodium bicarbonate buffer pH-7.4 and added to ~6.0 Log10 CFUs in an ultralow binding 96 well U-bottom plate. The plates were incubated at 37^oC overnight in moisture controlled container in an incubator. MIC was determined as the lowest concentration of agent that inhibits bacterial growth detected by naked eye. [0075] After the MIC assays, a fraction of sample from the wells were diluted in Mueller Hinton™ (MH) broth and plated on MH agar plates to enumerate the CFUs and determine the MIC and MBC of each PNA-CPP derivative against P. aeruginosa and A. baumannii isolates. The results are shown in Tables 5 and 6.

Example 4 Hemolysis Activity of PPNA Compounds in Red Blood Cells (RBCs) [0076] Hemolysis activity of PPNA compounds determined in human, rat, and mouse RBCs. Human, rat, and mouse RBCs purchased from commercial vendors. RBCs were washed in 10 mM HEPES buffer. PPNA compounds were prepared in 10mM sodium bicarbonate buffer pH 7.4. One hundred microliter of RBCs were exposed to different concentrations of PPNA compounds and incubated for two hours at 37^oC in Eppendorf tubes. Triton-X^TM (0.1%) and sodium bicarbonate buffer used as the positive and negative controls. Eppendorf tubes were centrifuged at 5000g, supernatants collected, and optical density determined at 530nm in a spectrophotometer. Table 7 represents the percentage of lysis of RBC at 500 µg/mL. Table 7: Hemolysis activity of PPNA compounds in RBCs

Example 5 Cytotoxic Effect of PPNA compounds in cell lines [0077] Cytotoxic effect of PPNA compounds were evaluated in five different cell lines (HepG2, human primary keratinocytes, human renal epithelial cells (HREC), mouse fibroblast cells, and rat fibroblast cells). Cells were purchased from ATCC and grown in recommended culture medium in presence or absence of serum. Cells (2x10⁴/well) were plated in 96 well flat bottom tissue culture plates. PPNA compounds were prepared in 10mM sodium bicarbonate buffer and different concentrations were added on to the plates. Mitomycin C and 1% Triton X^TM used as controls. Promega CellTiter-Glo^TM Luminescent Cell Viability Assay kit used to determine the cytotoxicity of PPNA compounds in five different cells. Reagents for the cell viability assay were prepared following manufacturer’s instruction and the luminescence of samples determined in Clariostar™ Plus spectrophotometer. Tables 8 and 9 represent the percentage cell viability in presence of PPNA compounds at 100 µg/mL. Table 8: Cell Viability at 100 µg/mL of P. aeruginosa PPNAs

Table 9: Cell Viability at 100 µg/mL of A. baumannii PPNAs

Example 6 Single Intravenous Dose Administration to Determine the Maximum Tolerability Dose (MTD) of Drugs in Mice. [0078] Ascending intravenous (IV) and intraperitoneal (IP), or continuous infusion subcutaneous dose study was performed to determine tolerability of the drug in infected mice. The animals will be made neutropenic by administration of cyclophosphamide (Cytoxan^TM) on Days (-4: 150 mg/kg) and (-1: 100 mg/kg) via IP route. Mice were infected intramuscularly on Day-0 with of P. aeruginosa (ATCC 39324) and A. baumannii (UNT191.1) isolates. At the time of PPNA 23X-PEG6-CPP2 and PPNA 25X- PEG6-CPP2 administration (2 hours post-infection), mean CFU titers of P.aeruginosa were 4.29log₁₀ and 4.75log₁₀ CFUs, respectively. Similarly, at the time of PPNA 39- PEG6-HomoArg administration (2 hours post-infection), mean CFU titers of A. baumannii were 8.20log10 CFUs. [0079] Initially, compounds PPNA 23X-PEG6-CPP2, PPNA 23X-PEG6-CPP2 and PPNA 39-PEG6-HomoArg were formulated in 10mM NaHCO₃ buffer and administered 2 hours post-infection at IV, IP, and SC route in infected mice. Compounds PPNA 23X- PEG6-CPP2, PPNA 23X-PEG6-CPP2 and PPNA 39-PEG6-HomoArg were administered 10 mg/mL three times within 24 hours in bolus doses for IV and IP routes. However, compounds were administered 30 mg/mL subcutaneously via Alzet^TM pumps for 24 hours continuous infusion. Alzet^TM pumps were implanted subcutaneously in individual mouse by surgical procedure. The MTD of PPNA 23X-PEG6-CPP2, PPNA 25X-PEG6-CPP2 and PPNA 39-PEG6-HomoArg are shown in Table 10. Higher concentration more than MTD via IV and IP route shown adverse effect and death in mice within 45 mins of administration of the drug. However, the exact MTD for SC route of administration is not determined. Table 10: Maximum Tolerability Dose (MTD) of PPNA 23X-PEG6-CPP2, PPNA 25X-PEG6- CPP2 and PPNA 39-PEG6-HomoArg

Example 7 Murine Thigh Infection Model to Determine the Antimicrobial Efficacy of Drugs Against P. aeruginosa and A. baumannii Infection [0080] Female 5-6 week old CD-1 (18-22 gm) were used in this study. Mice were quarantined for 48 hours before use and housed in groups of 5 with free access to food and water during the study. [0081] The animals were made neutropenic by administration of cyclophosphamide on Days (-4) and (-1). On Days (-4) 150 mg/kg of cyclophosphamide administered by intraperitoneal route and 100 mg/kg was administered on Days -1. Days listed are referenced from the date of infection (study day-Day 0). Example 8 Efficacy of PPNA 23X-PEG6-CPP2 and PPNA 25X-PEG6-CPP2 in the Thigh Infection Model with P. aeruginosa (CDC-FDA-AR-BANK #0233): [0082] On Day 0, animals were inoculated intramuscularly (0.1ml/thigh) with 6.01log₁₀ and 5.75log10 CFU/mouse of P. aeruginosa (CDC-FDA-AR-BANK #0233) into right thighs for testing of PPNA 23X-PEG6-CPP2 and PPNA 25X-PG6-CPP2 compounds, respectively. [0083] Mice were euthanized by CO₂ inhalation and thighs were removed, after the end of the study and placed in 2 ml of sterile PBS, homogenized, serially diluted and plated to determine the CFU counts. Plates were incubated 18-24 hours and CFUs were counted. Colony were counted and the number of colonies is converted to CFU/thigh by multiplying the number of colonies by the volume of the thigh homogenate spotted and the dilution at which the colonies were counted (5-50 colonies/spot). CFU count data were transformed into log10 CFU/thigh for calculation of means and standard deviations. [0084] The dosing regimen, CFU counts of and mean bacterial thigh titers for PPNA 23X-PEG6-CPP2 and PPNA 25X-PEG6-CPP2 treated groups of mice are shown in Tables 11 and 12.

[0085] The efficacy of PPNA 23X-PEG6-CPP2 (PPNA 23X) exhibited for all dose groups following both IV or IP dosing and continuous SC infusion was determined to be statistically significant when compared to the 1 hr or 24 hr untreated controls (p<0.015) (Figure 17).

[0086] The efficacy exhibited by PPNA 25X-PEG6-CPP2 (PPNA 25X) for all dose groups following both IP dosing and continuous SC infusion at 300 mg/kg was determined to be statistically significant when compared to the 1hr or 24 hr untreated controls (p<0.0001) (Figure 18). Example 9 Efficacy of PPNA 39 in the Thigh Infection Model with A. baumannii UNT191-1 [0087] Mice were inoculated IM with 5.27 log₁₀ CFU of A. baumannii UNT191-1 resulting in mean bacterial thigh titers of 6.28 log₁₀ CFU at 1 hour post-infection (start of treatment). The efficacy exhibited by PPNA 39 for all IV dose groups was determined to be statistically significant when compared to both the 1 and 24 hr untreated controls (p<0.0001). The dosing regimen, CFU counts of and mean bacterial thigh titers for PPNA 39-PEG6-HomoArg treated groups of mice are shown in Table 13. Table 13: The Dosing Regimen, CFU Counts of and Mean Bacterial Thigh Titers for PPNA 39- PEG6-HomoArg Treated Groups of Mice

Claims

WHAT IS CLAIMED IS: 1. A compound as depicted in the following table

wherein L_x is a linker, X is -NH-(CH₂)₅C(O)-, R is Arg, hR is homoArg, and x is 1, 2 or 3, or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein Lx is -NH-PEG-CH2CH2C(O)-, wherein PEG is polyethylene glycol of 2-10 repeating units; -NH-PEG-CH2C(O)-, wherein PEG is of 2- 10 repeating units; or -NH-PEG-CH₂CH₂NHC(O)CH₂CH₂C(O)-, wherein PEG is of 2-10 repeating units.

3. The compound of claim 1, wherein L_x comprises a PEG linker of 6 repeating units.

4. The compound of claim 1, wherein the polynucleotide of the compound comprises a modified backbone.

5. The compound of claim 1, wherein the compound is depicted in Fig 2, or a pharmaceutically acceptable salt thereof.

6. The compound of claim 1, wherein the compound is depicted in Fig 3, or a pharmaceutically acceptable salt thereof.

7. The compound of claim 1, wherein the compound is depicted in Fig 4, or a pharmaceutically acceptable salt thereof.

8. The compound of claim 1, wherein the compound is depicted in Fig 5, or a pharmaceutically acceptable salt thereof.

9. The compound of claim 1, wherein the compound is depicted in Fig 6, or a pharmaceutically acceptable salt thereof.

10. The compound of claim 1, wherein the compound is depicted in Fig 7, or a pharmaceutically acceptable salt thereof.

11. The compound of claim 1, wherein the compound is depicted in Fig 8, or a pharmaceutically acceptable salt thereof.

12. The compound of claim 1, wherein the compound is depicted in Fig 9, or a pharmaceutically acceptable salt thereof.

13. The compound of claim 1, wherein the compound is depicted in Fig 10, or a pharmaceutically acceptable salt thereof.

14. The compound of claim 1, wherein the compound is depicted in Fig 11, or a pharmaceutically acceptable salt thereof.

15. The compound of claim 1, wherein the compound is depicted in Fig 12, or a pharmaceutically acceptable salt thereof.

16. The compound of claim 1, wherein the compound is depicted in Fig 13, or a pharmaceutically acceptable salt thereof.

17. The compound of claim 1, wherein the compound is depicted in Fig 14, or a pharmaceutically acceptable salt thereof.

18. The compound of claim 1, wherein the compound is depicted in Fig 15, or a pharmaceutically acceptable salt thereof.

19. A pharmaceutical composition comprising the compound of any one of claims 1 to 18 and a pharmaceutically acceptable carrier.

20. A method of treating a bacterial infection, comprising administering to an animal in need thereof an effective amount of the compound of any one of claims 1 to 18 or pharmaceutical composition of claim 19.

21. The method of 20, wherein the bacteria is Pseudomonas and/or Acinetobacter.

22. The method of claim 21, wherein the bacteria is Pseudomonas aeruginosa.

23. The method of claim 21, wherein the bacteria is Acinetobacter baumannii.

24. The method of any one of claims 20-23, wherein the animal has hospital/ventilator associated pneumonia.

25. The method of any one of claims 20-23, wherein the animal has sepsis.

26. The method of any one of claims 20-23, wherein the animal has a urinary tract infection.

27. The method of any one of claims 20-23, wherein the animal has soft tissue-associated infection.

28. The method of any one of claims 20-23, wherein the animal has hardware prosthetic- associated infection.

29. The method of any one of claims 20-28, wherein the treatment route comprises intravenous, subcutaneous or oral administration.

30. The method of any one of claims 20-29, wherein the animal is a human.