Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/08—Tripeptides
  • C07K5/0819—Tripeptides with the first amino acid being acidic
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
  • A61K38/08—Peptides having 5 to 11 amino acids
  • A61K38/085—Angiotensins
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/04—Antibacterial agents
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/08—Tripeptides
  • C07K5/0821—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/10—Tetrapeptides
  • C07K5/1002—Tetrapeptides with the first amino acid being neutral
  • C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
  • C07K5/101—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/10—Tetrapeptides
  • C07K5/1021—Tetrapeptides with the first amino acid being acidic
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
  • C07K7/04—Linear peptides containing only normal peptide links
  • C07K7/14—Angiotensins: Related peptides
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
  • C12N2501/30—Hormones
  • C12N2501/32—Angiotensins [AT], angiotensinogen

Definitions

• the predominant pathogen is Escherichia coli, although other gram-negative bacteria such as the Klebsiella-Enterobacter-Serratia group and Pseudomona, and gram positive microbes such as Staphylococcus, can be the causative pathogen.
• Escherichia coli gram-negative bacteria
• Staphylococcus gram positive microbes
• the genitourinary tract, gastrointestinal tract and respiratory tract are the most frequent sources of sepsis. Other common foci are wound, burn, and pelvic infections and infected intravenous catheters.
• Gram negative sepsis is a disease syndrome resulting from the systemic invasion of gram negative rods and subsequent endotoxemia.
• the severity of the disease ranges from a transient, self-limiting episode of bacteremia to a fulminant, life-threatening illness often complicated by organ failure and shock.
• the disease is often the result of invasion from a localized infection site, or may result from trauma, wounds, ulcerations or gastrointestinal obstructions.
• the symptoms of gram negative sepsis include fever, chills, pulmonary failure and septic shock (severe hypotension).
• Gram negative infections are particularly common among patients receiving anti-cancer chemotherapy and immunosuppressive treatment.
• U.S. Patent No. 5,698,198 Infections in such immuno-compromised hosts characteristically exhibit resistance to many antibiotics, or develop resistance over the long course of the infection, making conventional treatment difficult.
• the ever increasing use of cytotoxic and immunosuppressive therapy and the natural selection for drug resistant bacteria by the extensive use of antibiotics have contributed to gram negative bacteria evolving into pathogens of major clinical significance.
• Septic shock is a major cause of death in intensive care units. It is estimated that over 700,000 patients become susceptible to septic shock-causing bacterial infections each year in the United States alone. Of these, 160,000 actually develop septic shock, resulting in 50,000 deaths annually.
• the present invention provides methods and kits for treating or preventing infection comprising the administration of an amount effective for treating or preventing an infection of angiotensinogen, angiotensin I (Al), Al analogues, Al fragments and analogues thereof, angiotensin II (All), All analogues, All fragments or analogues thereof or All AT 2 type 2 receptor agonists.
• the present invention provides improved methods and pharmaceutical compositions for antibiotic therapy, wherein the improvement comprises the administration of an amount effective for treating a bacterial infection
• angiotensinogen Al, Al analogues, Al fragments and analogues thereof, All, All analogues, All fragments or analogues thereof or All AT 2 type 2 receptor agonists.
• Figure 1 is a graph showing the effect of All on host resistance to bacterial peritonitis, based on the percentage of peritoneal sites without abscesses associated with infection.
• Figure 2 is a graph showing the effect of All on host resistance to bacterial peritonitis, based on the mean overall abscess score.
• Figure 3 is a graph showing the effect of All administration on abscess score (mean score).
• Figure 4 is a graph showing the effect of All administration on abscess score (rank order analysis).
• Figure 5 is a graph showing the effect of All administration on abscess incidence.
• Figure 6 is a graph showing the effect of All with and without Ofloxacin on abscess formation (mean score).
• Figure 7 is a graph showing the effect of All with and without Ofloxacin on abscess formation (rank order).
• Figure 8 is a graph showing the effect of All with and without Ofloxacin on abscess free sites (mean score).
• Figure 9 is a graph showing a comparison of All, AII(l-7) and Neupogen in a rat peritonitis model (mean abscess score).
• Figure 10 is a graph showing a comparison of All, AII(l-7) and Neupogen in a rat peritonitis model (rank order).
• Figure 11 is a graph showing a comparison of All, AII(l-7) and Neupogen in a rat peritonitis model (% abscess free).
• Figure 12 is a graph showing a comparison of AII(l-7) analogues in the infection model (mean abscess score).
• Figure 13 is a graph showing a comparison of AII(l-7) analogues in the infection model (rank order).
• Figure 14 is a graph showing a comparison of AII(l-7) analogues in the infection model (% abscess free).
• Infections are broadly defined to mean situations when the invasion of a host by an agent is associated with the clinical manifestations of infection including, but not limited to, at least one of the following: abnormal temperature, increased heart rate, abnormal respiratory rate, abnormal white blood cell count, fatigue, chills, muscle ache, pain, dizziness, dehydration, vomiting, diarrhea, and organ dysfunction.
• infections may be bacterial, viral, or parasitic in nature.
• All in an amount which is sufficient for said increase.
• the application of All to wound tissue significantly increases the rate of wound healing, leading to a more rapid re-epithelialization and tissue repair.
• the term All refers to an octapeptide present in humans and other species having the sequence Asp-Arg-Val-Tyr-Ile-His- Pro-Phe [SEQ ID NO:l].
• the biological formation of angiotensin is initiated by the action of renin on the plasma substrate angiotensinogen (Clouston et al., Genomics 2:240-248 (1988); Kageyama et al, Biochemistry 23:3603-3609; Ohkubo et al., Proc. Natl. Acad. Sci.
• angiotensin I (Al) which is converted to All by the angiotensin converting enzyme (ACE) which removes the C-terminal His-Leu residues from Al [SEQ ID NO: 37]. All is a known pressor agent and is commercially available.
• mice Treatment of mice with an ACE inhibitor, which acts to prevent the formation of All from its precursor angiotensin I, after thermal injury resulted in greater survival and decreased bacterial translocation compared to controls. (Gennari et al., Shock 6:95-100 (1996))
• 7 receptor agonists would be useful for the treatment and prevention of bacterial, viral, or parasitic infections, or that angiotensinogen, Al, All, Al or All analogues or fragments or All AT 2 type 2 receptor agonists would be useful as an improvement for antibiotic therapy.
• a peptide agonist selective for the AT2 receptor (All has 100 times higher affinity for AT2 than ATI) has been identified.
• This peptide is p- aminophenylalanine 6-AII ["(p-NH 2 -Phe) 6- All)"], Asp-Arg-Val-Tyr-Ile-Xaa-Pro- Phe [SEQ ID NO.36] wherein Xaa is p-NH 2 -Phe (Speth and Kim, BBRC 169:997- 1006 (1990).
• This peptide gave binding characteristics comparable to AT2 antagonists in the experimental models tested (Catalioto, et al., Eur. J. Pharmacol. 256:93-97 (1994); Bryson, et al., Ewr. J. Pharmacol. 225:119-127 (1992).
• AII(l-7) All residues 1-7) or other fragments of All to evaluate their activity.
• AII(l-7) elicits some, but not the full range of effects elicited by AIL Pfeilschifter, et al., Eur. J. Pharmacol. 225:57-62
• a preferred class of AT2 agonists for use in accordance with the present invention comprises angiotensinogen, angiotensin I (Al), Al analogues, Al fragments and analogues thereof, All, All analogues, All fragments or analogues thereof or All AT 2 type 2 receptor agonists having p-NH- Phe in a position corresponding to a position 6 of AIL
• various nonpeptidic agents e.g., peptidomimetics
• having the requisite AT2 agonist activity are further contemplated for use in accordance with the present invention.
• the active All analogues, fragments of All and analogues thereof of particular interest in accordance with the present invention comprise a sequence consisting of at least three contiguous amino acids of groups R3R 8 in the sequence of general formula I
• R 1 and R 2 together form a group of formula X-R A -R B -, wherein X is H or a one to three peptide group,
• R A is suitably selected from Asp, Glu, Asn, Acpc (1- aminocyclopentane carboxylic acid), Ala, Me 2 Gly, Pro, Bet, Glu(NH 2 ), Gly, Asp(NH 2 ) and Sue
• R B is suitably selected from Arg, Lys, Ala, Orn, Ser(Ac), Sar, D-Arg and D-Lys;
• R 3 is selected from the group consisting of Val, Ala, Leu, Lys, norLeu, lie, Gly, Pro, Aib, Acpc and Tyr;
• R 4 is selected from the group consisting of Tyr, Tyr(PO ) 2 , Thr, Ser, Ala, homo Ser and azaTyr;
• R 5 is selected from the group consisting of He, Ala, Leu, norLeu, Val
• R 6 is His, Arg or 6-NH 2 -Phe
• R is Pro or Ala
• R is selected from the group consisting of Phe, Phe(Br), He and Tyr, excluding sequences including R 4 as a terminal Tyr group.
• Compounds falling within the category of AT2 agonists useful in the practice of the invention include the All analogues set forth above subject to the restriction that R 6 is p-NH 2 -Phe.
• R and R are Asp-Arg, Asp-Lys, Glu-Arg and Glu-Lys.
• Particularly preferred embodiments of this class include the following: All, AIII or AII(2-8), Arg-Val-Tyr-Ile-His-Pro-Phe [SEQ ID NO:2]; AII(3-8), also known as desl-AIII or AIV, Val-Tyr-Ile-His-Pro-Phe [SEQ ID NO:
• AII(l-7) Asp-Arg-Val-Tyr-Ile-His-Pro ⁇ SEQ ID NO:4]; AII(2-7).
• Arg-Val- Tyr-Ile-His-Pro [SEQ ID NO:5]; AII(3-7), Val-Tyr-Ile-His-Pro [SEQ ID NO:6]; AII(5-8), Ile-His-Pro-Phe [SEQ ID NO:7]; AII(l-6), Asp-Arg-Val-Tyr-Ile-His [SEQ ID NO:8]; AII(l-5), Asp-Arg- Val-Tyr-Ile [SEQ ID NO:9]; AII(l-4), Asp-Arg-Val- Tyr [SEQ ID NO:10]; and AII(l-3), Asp-Arg-Val [SEQ ID NO:l 1].
• Other preferred embodiments include: Arg-norLeu-Tyr-Ile-His-Pro-Phe [SEQ
• Still another preferred embodiment encompassed within the scope of the invention is a peptide having the sequence
• the active compounds of the present invention are selected from those comprising the following general formula: Asp-Arg-Rl-R2-Ile-His-Pro-R3, wherein
• Rl is selected from the group consisting of Val, Pro, Lys, Norleu, and Leu;
• R2 is selected from the group consisting of Ala, Tyr, and Tyr(PO 3 ) 2 ;
• R3 is Phe or is absent.
• R 2 is selected from the group consisting of H, Arg, Lys, Ala, Orn, Ser(Ac), Sar, D-Arg and D-Lys;
• R 3 is selected from the group consisting of Val, Ala, Leu, norLeu, He, Gly, Pro, Aib, Acpc and Tyr;
• R 4 is selected from the group consisting of Tyr, Tyr(PO 3 ) 2 , Thr, Ser, homoSer and azaTyr;
• R 5 is selected from the group consisting of He, Ala, Leu, norLeu, Val and Gly;
• R 6 is His, Arg or 6-NH 2 -Phe;
• R 7 is Pro or Ala
• R is selected from the group consisting of Phe, Phe(Br), He and Tyr.
• R 2 , R 3 and R 5 are as previously defined.
• Particularly preferred is angiotensin III of the formula Arg- Val-Tyr-Ile-His-Pro-Phe [SEQ ID NO:2].
• Other preferred compounds include peptides having the structures Arg-Val-Tyr-Gly-His- Pro-Phe [SEQ ID NO: 17] and Arg-Val-Tyr-Ala-His-Pro-Phe [SEQ ID NO: 18].
• the fragment AII(4-8) was ineffective in repeated tests; this is believed to be due to the exposed tyrosine on the N-terminus.
• Bet 1 carboxy-N,N,N-trimethylmethanaminium hydroxide inner salt (betaine)
• R 2 Appropriate side chains on the amino acid in position R 2 may contribute to affinity of the compounds for target receptors and/or play an important role in the conformation of the peptide. For this reason, Arg and Lys are particularly preferred as R 2 .
• R may be involved in the formation of linear or nonlinear hydrogen bonds with R 5 (in the gamma turn model) or R 6 (in the beta turn model).
• R 3 would also participate in the first turn in a beta antiparallel structure (which has also been proposed as a possible structure).
• beta and gamma branching are equally effective in this position.
• a single hydrogen bond may be sufficient to maintain a relatively stable conformation.
• R may suitably be selected from Val, Ala, Leu, norLeu, He, Gly, Pro, Aib, Acpc and Tyr.
• Lys has also been found to be effective at R .
• R 4 is
• Tyr 13 preferably selected from Tyr, Thr, Tyr (PO 3 ) 2 , homoSer, Ser and azaTyr.
• Tyr is particularly preferred as it may form a hydrogen bond with the receptor site capable of accepting a hydrogen from the phenolic hydroxyl (Regoli, et al. (1974), supra).
• Ala has also been found to be effective at R 4 .
• Gly is suitable in position R 5 , it is preferred that the amino acid in this position be selected from He, Ala, Leu, norLeu, Gly and Val.
• R 6 is His, Arg or 6-NH 2 -Phe.
• the unique properties of the imidazole ring of histidine e.g., ionization at physiological pH, ability to act as proton donor or acceptor, aromatic character) are believed to contribute to its particular utility as R 6 .
• conformational models suggest that His may participate in hydrogen bond formation (in the beta model) or in the second turn of the antiparallel structure by influencing the orientation of R 7 .
• R 7 should be Pro in order to provide the most desirable orientation of R .
• both a hydrophobic ring and an anionic carboxyl terminal appear to be particularly useful in binding of the analogues of interest to receptors; therefore, Tyr and especially Phe are preferred for purposes of the present invention.
• Analogues of particular interest include the following:
• Analogue 16 Asp-Arg-Val-Tyr-norLeu-His-Pro-Phe SEQ ID NO: 34
• Analogue 17 Asp-Arg-Val-homoSer-Tyr-Ile-His-Pro-Phe SEQ ID NO: 35
• polypeptides of the instant invention may be produced by any standard method, including but not limited to recombinant DNA technology and conventional synthetic methods including, but not limited to, those set forth in J. M. Stewart and
• peptides are synthesized according to standard solid-phase methodologies, such as may be performed on an Applied Biosystems Model 430 A peptide synthesizer (Applied Biosystems, Foster City, Calif), according to manufacturer's instructions. Other methods of synthesizing peptides or peptidomimetics, either by solid phase methodologies or in liquid phase, are well known to those skilled in the art.
• the present invention provides methods and kits for treating and preventing infections in a mammal comprising administering to the mammal an amount effective to treat or prevent an infection of angiotensinogen, angiotensin I (Al), Al analogues, Al fragments and analogues thereof, angiotensin II (All), AH analogues, AH fragments or analogues thereof or AH AT 2 type 2 receptor agonists (the active agents).
• the invention is appropriate for the treatment and prevention of all types of infection, including but not limited to septic shock, peritonitis, bacteremia, endotoxemia, and viral and parasitic infections.
• the methods of the invention are applicable to infections resulting from any condition, including but not limited to
• the active agents of the invention can be used alone or in a combination of active agents, or may be used in combination with other anti-infective agents, including but not limited to oflaxacin, granulocyte colony stimulating factors, gentamicin, augmentin or cephalosporins such as ceftazidime, amino-glycosides (such as amikacin, tobramycin, netilmicin, and gentamicin), related beta-lactam agents such as maxalactam, carbopenems such as imipenem, monobactam agents such as aztreonam; ampicillin and broad-spectrum penicillins, (e.g., penicillinase- resistant penicillins, ureidopenicillins or antipseudomonal penicillin or Augmentin) that are active against P.
• anti-infective agents including but not limited to oflaxacin, granulocyte colony stimulating factors, gentamicin, augmentin or cephalosporins such as cefta
• the present invention provides improved methods, compositions, and kits for antibiotic therapy, wherein the improvement comprises the administration of an amount effective for treating a bacterial infection of the active agents.
• the active agents may be administered by any suitable route, including orally, parentally, by inhalation spray, rectally, or topically in dosage unit formulations containing conventional pharmaceutically acceptable carriers, adjuvants, and vehicles.
• parenteral as used herein includes, subcutaneous, intravenous, intraarterial, intramuscular, intrasternal, intratendinous, intraspinal, intracranial, intrathoracic, infusion techniques or intraperitoneally.
• the active agents may be made up in a solid form (including granules, powders or suppositories) or in a liquid form (e.g., solutions, suspensions, or emulsions).
• the compounds of the invention may be applied in a variety of solutions. Suitable solutions for use in accordance with the invention are sterile, dissolve sufficient amounts of the peptide, and are not harmful for the proposed application. In this regard, the compounds of the present invention are very stable but are hydrolyzed by strong acids and bases.
• the compounds of the present invention are soluble in organic solvents and in aqueous solutions at pH 5-8.
• the active agents may be subjected to conventional pharmaceutical operations such as sterilization and/or may contain conventional adjuvants, such as preservatives, stabilizers, wetting agents, emulsifiers, buffers etc.
• the active agents are ordinarily combined with one or more adjuvants appropriate for the indicated route of administration.
• the compounds may be admixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acids, stearic acid, talc, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulphuric acids, acacia, gelatin, sodium alginate, polyvinylpyrrolidine, and/or polyvinyl alcohol, and tableted or encapsulated for conventional administration.
• the compounds of this invention may be dissolved in saline, water, polyethylene glycol, propylene glycol, carboxymethyl cellulose colloidal solutions, ethanol, com oil, peanut oil, cottonseed oil, sesame oil, tragacanth gum, and/or various buffers.
• Other adjuvants and modes of administration are well known in the pharmaceutical art.
• the carrier or diluent may include time delay material, such as glyceryl monostearate or glyceryl distearate alone or with a wax, or other materials well known in the art.
• Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin ⁇ e.g., liniments, lotions, ointments, creams, or pastes) and drops suitable for administration to the eye, ear, or nose.
• the dosage regimen for treating or preventing infections in a mammal with the active agents is based on a variety of factors, including the type of injury, the age, weight, sex, medical condition of the individual, the severity of the condition, the route of administration, and the particular compound employed. Thus, the dosage regimen may vary widely, but can be determined routinely by a physician using standard methods. Dosage levels of the order of between 0.1 ng/kg and 10 mg/kg body weight active agent are useful for all methods of use disclosed herein.
• the treatment regime will also vary depending on the infection being treated, based on a variety of factors, including the type of infection, the age, weight, sex, medical condition of the individual, the severity of the condition, the route of administration, and the particular compound employed.
• the active agents are administered to a mammal suffering from bacteremia for two weeks.
• the therapy is administered for between two and five times per day at dosages as described above.
• the active agent is administered subcutaneously or intraperitoneally.
• a suitable subcutaneous dose of active agent is preferably between about 0.1 ng/kg and about 10 mg/kg administered twice daily for a time sufficient to treat or prevent infections in a mammal.
• the concentration of active agent is between about 100 ng/kg body weight and about 10.0 mg/kg body weight.
• the active agent is administered subcutaneously or intraperitoneally.
• a suitable subcutaneous dose of active agent is preferably between about 0.1 ng/kg and about 10 mg/kg administered twice daily for a time sufficient to treat or prevent infections in a mammal.
• the concentration of active agent is between about 100 ng/kg body weight and about 10.0 mg/kg body weight.
• the active agent is administered subcutaneously or intraperitoneally.
• the active ingredient may comprise from
• 0.0001% to 10% w/w e.g., from 1% to 2% by weight of the formulation, although it may comprise as much as 10% w/w, but preferably not more than 5% w/w, and more preferably from 0.1% to 1% of the formulation.
• the active agent is administered topically.
• Suitable topical doses and active ingredient concentration in the formulation are as described for subcutaneous administration.
• kits for treating or preventing infection in a mammal wherein the kits comprise an effective amount of the active agents for treating or preventing infection in a mammal, and instructions for using the amount effective of active agent as a therapeutic.
• the kit further comprises a pharmaceutically acceptable carrier, such as those adjuvants described above.
• the kit further comprises a means for delivery of the active agent to a patient.
• Such devices include, but are not limited to syringes, matrical or micellar solutions, bandages, wound dressings, aerosol sprays, lipid foams, transdermal patches, topical administrative agents, polyethylene glycol polymers, carboxymethyl cellulose preparations, crystalloid preparations (e.g., saline, Ringer's lactate solution, phosphate-buffered saline, etc.), viscoelastics, polyethylene glycols, and polypropylene glycols.
• the means for delivery may either contain the effective
• the active agent 20 amount of the active agent, or may be separate from the active agents, which are then applied to the means for delivery at the time of use.
• the active agents can be administered alone, or may be combined with other anti-infective agents in combinatorial therapy.
• the anti- infective agent is selected from the group consisting of Ofloxacin and granulocyte colony stimulating factors.
• the method comprises pharmaceutical compositions for treating or preventing infections, comprising the active agents of the invention, an amount an amount effective to treat or prevent an infection of an anti-infective agent and a pharmaceutically acceptable carrier.
• the anti-infective agent is select4ed from the group consisting of
• the methods and kits of the present invention provide significant benefits for the treatment and prophylaxis of mammalian infections.
• the methods and kits of the present invention may be particularly valuable in the hospital setting, where potentially serious bacterial infections are common, and may also enable decreased reliance on high doses of antibiotics, which can lead to the development of antibiotic resistant bacteria.
• the following rat model of bacteremia was used in all subsequent examples. Forty-five female Sprague Dawley rats, weighing between 175 and 225 grams each, were used in the study. Fifteen of the rats were used to produce fecal material. The rats were housed in the University of Southern California vivarium on a 12:12 hour light/dark cycle and were quarantined at least two days prior to surgery. Food and water were available ad libitum except in the immediate postoperative period.
• the rats underwent a standardized procedure for laparotomy (intramuscular anesthesia with ketamine/rompum, shaving with animal clippers, betadine scrub, alcohol scrub). A 2 cm incision was then made on the midline. A double-walled gelatin capsule was placed on the right side of the abdomen through the incision. A polyethylene tube (PE 60) was sutured to the left sidewall and attached to the Alzot miniosmotic pump containing either saline, 10 ⁇ g/kg/day AH, or 100 ⁇ g/kg/day AIL The pump was then placed in a subcutaneous pocket. The abdominal wall and skin was then sutured closed using two layers of 4-0 Ethilon suture. Following surgery,
• the rats received analgesic for three days and were observed twice daily for signs of morbidity and mortality.
• Rats that died during the 11 day post-operative observation period were necropsied to confirm the presence of an acute bacterial infection.
• the rats that survived the initial acute infection were terminated on day 12 after surgery.
• Each rat was examined for odor upon opening and splenomegaly.
• four areas of the peritoneum were examined for abscess formation. These areas included the liver, abdominal wall, bowel and omentum.
• the abscesses were scored at each site as follows:
• the rat peritonitis model was generated as in Example 1. All (100 ⁇ g/kg/day) was given either: (1) subcutaneously (daily) three days before and after initiation of infection (SQ/SQ); (2) subcutaneously only after initiation of infection (SQ Post); (3) subcutaneously (daily) three days before and intraperitoneally after
• IP Post intraperitoneally via Alzet pump starting at the initiation of infection throughout the post-infection interval (Pump).
• the rat peritonitis model was generated as in Example 1. AH (100%)
• the rat peritonitis model was generated as in Example 1. AH (1-100 ⁇ g/kg/day) and AII(l-7) (1-100 ⁇ g/kg/day) treatment were compared with G-CSF treatment (Neupogen— Amgen, Thousand Oaks, CA) (0.1-10 ⁇ g/kg/day) for reduction in abscess size and occurrence.
• G-CSF treatment Neurogen— Amgen, Thousand Oaks, CA
• the experimental treatments were given by subcutaneous injection starting three days prior to initiation of infection and continued until the animals were euthanized.
• Example 5 AII(l-7) analogue effect on abscess formation
• the rat peritonitis model was generated as in Example 1. Subcutaneous injections with the peptides listed in Table 6 (10-100 ⁇ g/kg/day) were initiated three days prior to the initiation of infection and continued until the animals were euthanized.

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