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WO2024249553A1 - Molécules ciblant des tissus infectés et leurs procédés d'utilisation - Google Patents

Molécules ciblant des tissus infectés et leurs procédés d'utilisation Download PDF

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Publication number
WO2024249553A1
WO2024249553A1 PCT/US2024/031548 US2024031548W WO2024249553A1 WO 2024249553 A1 WO2024249553 A1 WO 2024249553A1 US 2024031548 W US2024031548 W US 2024031548W WO 2024249553 A1 WO2024249553 A1 WO 2024249553A1
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Prior art keywords
seq
forms
peptide
infected tissue
residues
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English (en)
Inventor
Tambet Teesalu
Erkki Ruoslahti
Gregory T. Robertson
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Tartu Ulikool (University of Tartu)
Colorado State University Research Foundation
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Tartu Ulikool (University of Tartu)
Colorado State University Research Foundation
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Publication of WO2024249553A1 publication Critical patent/WO2024249553A1/fr
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    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/03—Peptides having up to 20 amino acids in an undefined or only partially defined sequence; Derivatives thereof
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent

Definitions

  • the invention is generally directed to compositions and methods of peptides that target and bind to tissues infected by microbial pathogens , particularly peptides that selectively target and bind to tissues infected by bacteria, such as Mycobacterium tuberculosis.
  • Tuberculosis is a treatable disease caused by the bacterium Mycobacterium tuberculosis (Mtb ⁇ ).
  • TB is a difficult infection to treat, treatment of even uncomplicated, drug-susceptible tuberculosis (TB) is lengthy, requiring 6 to 9 months with a 40-year-old four-drug regimen for which extensive clinical drug resistance exists.
  • compositions that specifically recognize and home to tissues infected with microbial pathogens in vivo.
  • compositions including the infected tissue homing molecules in combination with one or more active agents, such as antimicrobial agents, and optionally a delivery vehicle are provided.
  • the compositions selectively target microbial pathogens and infected tissues in vivo and therefore selectively deliver the associated one or more active agents, such as antimicrobial agents, to microbial pathogens, such as cells of Mtb and/or infected tissues, such as tissues infected with Mtb.
  • the methods include administering to the subject a pharmaceutical formulation including (a) an infected tissue homing molecule; and (b) an antimicrobial agent, wherein the antimicrobial agent is conjugated or complexed with the infected tissue homing peptide, and wherein the antimicrobial agent is in an amount effective to treat or prevent one or more symptoms of the infection in the subject.
  • Methods for identification of an infection by a microbial pathogen in a subject including administering to the subject a pharmaceutical formulation including (a) an infected tissue homing molecule; and (b) an imaging or diagnostic agent, wherein the imaging or diagnostic agent is in an amount effective to bind to infected tissue and/or pathogens and label or otherwise identify the infection in the subject.
  • a pharmaceutical formulation including (a) an infected tissue homing molecule; and (b) an imaging or diagnostic agent, wherein the imaging or diagnostic agent is in an amount effective to bind to infected tissue and/or pathogens and label or otherwise identify the infection in the subject.
  • infected tissue homing molecule is or includes a peptide.
  • Exemplary infected tissuehoming peptides include a peptide having an amino acid sequence of any one or more of SEQ ID NOs:l-37, or a functional variant thereof having at least 70% sequence identity to any one of SEQ ID NOs: 1-37.
  • the infected tissue homing molecule is a peptide having an amino acid sequence PPRRGLIKLKTS (SEQ ID NO: 1).
  • the infected tissue homing molecule is a function variant of the peptide of SEQ ID NO: 1 , for example, having at least 70% sequence identity to PPRRGLIKLKTS (SEQ ID NO: 1).
  • the infected tissue homing molecule is a peptide that consists of PPRRGLIKLKTS (SEQ ID NO: 1).
  • the infected tissue homing molecule is or includes a peptide having an amino acid sequence AGRGRLVR (SEQ ID NO:2).
  • the infected tissue homing molecule is a function variant of the peptide of SEQ ID NO:2, for example, having at least 70% sequence identity to AGRGRLVR (SEQ ID NO:2).
  • the infected tissue homing molecule is a peptide that consists of AGRGRLVR (SEQ ID NOG).
  • the infected tissue homing molecule is a peptide having an amino acid sequence GRP ARP AR (SEQ ID NO:3).
  • the infected tissue homing molecule is a function variant of the peptide of SEQ ID NO:3, for example, having at least 70% sequence identity to GRP ARP AR (SEQ ID NOG).
  • the infected tissue homing molecule is a peptide that consists of GRP ARP AR (SEQ ID NOG).
  • the infected tissue homing molecule is a peptide having an amino acid sequence CKRDLSRRC (SEQ ID NOG).
  • the infected tissue homing molecule is a function variant of the peptide of SEQ ID NOG, for example, having at least 70% sequence identity to CKRDLSRRC (SEQ ID NOG).
  • the infected tissue homing molecule is a peptide that consists of CKRDLSRRC (SEQ ID NOG).
  • the infected tissue homing molecule is a peptide having an amino acid sequence AKRGARSTA (SEQ ID NOG).
  • the infected tissue homing molecule is a function variant of the peptide of SEQ ID NOG, for example, having at least 70% sequence identity to AKRGARSTA (SEQ ID NOG).
  • the infected tissue homing molecule is a peptide that consists of AKRGARSTA (SEQ ID NOG).
  • the methods administer a formulation that further includes one or more infected tissue homing molecules, such as polypeptides or other amino acid sequences, for example, whereby the polypeptides or other amino acid sequences are contiguous with the amino (NH) or carboxyl (COOH) terminus of the infected tissue homing peptide.
  • the methods administer a formulation that further includes
  • 45652286 one or more additional molecules selected from the group including a carbohydrate, a lipid, a small organic molecule, a polymer, and a nucleic acid.
  • the methods administer a formulation including an antimicrobial agent selected from bedaquiline, pretomanid, Rinzolid, delamanid, isoniazid, vancomycin, rifampin, rifabutin, pyrazinamide, ethambutol, streptomycin, amikacin, kanamycin, capreomycin, ofloxacin, levofloxacin, moxifloxacin, gatifloxacin, ethionamide, aminosalicylic acid, and cycloserine.
  • the antimicrobial is vancomycin.
  • the antimicrobial agent is attached to the peptide via a linker, such as a cleavable linker.
  • the methods administer a formulation including a delivery vehicle.
  • An exemplary delivery vehicle is a particle, such as a microparticle or a nanoparticle.
  • the delivery vehicle is selected from a citric acid silver nanoparticle, a liposome, a micelle, a viral capsid, a polymeric particle, a dendrimer, a porous silicon particle, a metal particle, and an iron oxide nanoparticle.
  • the delivery vehicle is a citric acid nanoparticle.
  • the methods include administering a formulation including a particle, the infected tissue homing molecule is conjugated to, or dispersed within the outer surface of the particle.
  • the infected tissue homing molecule is attached to the surface of the particle via a linker, such as a cleavable linker.
  • the infected tissue-targeting molecule is attached to the surface of the particle via a polyethylene glycol (PEG) linker.
  • the PEG has a molecular weight of between about 1,000 Da and about 10,000 Da such as about 5,000 Da.
  • the antimicrobial agent is encapsulated or embedded within one or more cavities and/or pores within the particle.
  • the particle has a mean hydrodynamic diameter of between about 40 nm and about 80 nm, inclusive, optionally wherein the particle has a mean diameter of about 60 nm.
  • the particle does not include another targeting or homing agent.
  • the particle is conjugated to between one and about 10,000 molecules of the infected tissue homing molecule.
  • the formulation is in a form selected from the group including a solution, a dry powder, micelles, colloids, nanodroplets, nanostructured hydrogel, nanocrystals, and a nanosuspension.
  • the microbial pathogen is one or more Mycobacterium sp. selected from the group including M. tuberculosis, M. africanum, M. canetti, M. bovis, M. caprae, M. microti, M. pinnipedii, M. mungi, M. leprae, M. avium, M. ulcerans, M. xenopi, M. kansasii, M. abscessus, M. chelonae, M. fortuitum and M. orygis.
  • the group including M. tuberculosis, M. africanum, M. canetti, M. bovis, M. caprae, M. microti, M. pinnipedii, M. mungi, M. leprae, M. avium, M. ulcerans, M. xenopi, M. kansasii, M. abscessus, M. chelonae, M. fortuitum
  • the 45652286 microbial pathogen is M. tuberculosis.
  • the microbial pathogen is an antibiotic resistant strain that is resistant to one or more antibiotics when administered in vivo alone at a safe dose.
  • the microbial pathogen is a Mycobacterium sp. that is an antibiotic resistant strain that is resistant to one or more antibiotics when administered in vivo alone at a safe dose.
  • antibiotics selected from the group including bedaquiline, pretomanid, Rinzolid, delamanid, isoniazid, vancomycin, rifampin, rifabutin, pyrazinamide, ethambutol, streptomycin, amikacin, kanamycin, capreomycin, ofloxacin, levofloxacin, moxifloxacin, gatifloxacin, ethionamide, aminosalicylic acid, and cycloserine, either alone (i.e., monoresistant) or in combination (polyresistant including multi-drug and Extensively-drug resistant forms of TB).
  • antibiotics selected from the group including bedaquiline, pretomanid, Rinzolid, delamanid, isoniazid, vancomycin, rifampin, rifabutin, pyrazinamide, ethambutol, streptomycin, amikacin, kanamycin, capreo
  • the antibiotic resistant strain is susceptible to the same antibiotic when administered conjugated with the infected tissue homing peptide.
  • the subject has been diagnosed as having, or as being at risk of having tuberculosis (TB) selected from pulmonary TB, CNS TB, pericardial TB, pleural TB, Lymphadenitis, Abdominal TB, skeletal TB, genitourinary TB and miliary TB.
  • the methods are effective to reduce, eliminate, or prevent one or more symptoms of Mtb infection selected from the group including fever, chest pain, wheezing or difficulty breathing, cough, fatigue, and unintentional weight loss.
  • the pharmaceutical formulation is administered to the subject via a route selected from oral administration, intramuscular injection, intravenous injection, sub-cutaneous injection, and intra-articular injection.
  • the effective amount of the antimicrobial agent administered in the formulation is less than the amount of the same therapeutic agent that is required to be effective when administered alone.
  • Dosage forms for treatment of an infection by a microbial pathogen are also described.
  • Dosage forms for treatment of an infection by Mycobacterium tuberculosis (Mtb) in a subject in need thereof are also described.
  • the dosage form includes (a) an infected tissue homing molecule including an amino acid sequence selected from SEQ ID NOs:l-37; and (b) an antimicrobial agent, whereby the antimicrobial agent is in an amount effective to treat or prevent one or more symptoms of the infection in the subject.
  • the effective amount of therapeutic agent administered in the formulation is less than the amount of the same therapeutic agent that is required to be effective when administered alone.
  • Figures 1A-1T are photo-micrograph images showing distribution of receptors of the candidate homing peptides in the TB-infected necrotic and cellular lesions. Images show TB-infected tissue sections stained with antibodies to identify the location of receptors in infected cellular lesions (Figs. 1A-1J) and infected necrotic lesions (Figs. 1K- 1P), respectively. Fluorescence immunostaining corresponds with presence of receptors including p32 (Figs. 1A, IK); RXRB (Figs. IB, IL); RelB (Figs. 1C, IM); Laminin (Figs. ID, IN); FN-EDB (Figs. IE, IO); HAS3 (Figs. IF, IP); TNC-C (Figs. 1G, IQ); NRP-1 (Figs. 1H, 1R); Fibrin (Figs. II, IS); and Alpha-v integrin (Figs. 1J, IT), respectively.
  • Figures 2A-2B are graphs of data from in vivo play-off experiments in TB- infected or healthy mice.
  • Fig. 2A shows the ratio of each of the targeting peptides (x-axis) to a control peptide (y-axis; 0 tolOO fold over G7 control phage) in each of TB-infected lung (dark bars) and non-infected lung (light bars) tissue, respectively
  • Fig. 2B shows the ratio of peptides (y-axis; 0-10 infected/noninfected lung phage representation) of each of the targeting peptides (x-axis), respectively.
  • FIGS 3A-3C are diagrams depicting peptide-functionalized silver nanoparticles (AgNP).
  • Fig. 3A is a graph of Absorbance [a.u.] over Wavelength [nm], with a photomicrograph of the particles also depicted. Scale bar is 200 nm;
  • Fig. 3B is a bar graph of Volume [%] over Hydro-dynamic Diameter [nm];
  • Fig. 3C is a diagram showing specifications of citric AgNPs used in the homing study, showing the silver (Ag) core covered with NeutrAvidin-PEG(5K)-thiols (boxes with lines) and Iipoic-PEG(1K)-NH2 (lines), respectively.
  • Targeting peptides are attached to NeutrAvidin and the payload is conjugated to the free NH2 groups of the lipoic-PEG(lK)-linkers.
  • Figure 4 is a flow chart depicting the experimental schema for in vivo homing studies using peptide-functionalized silver nanoparticles in Mycobacterium tuberculosis infected C3HeB/FeJ mice.
  • Figures 5A-5B are photo-micrograph images showing staining of infected tissue with FAM-labelled PL1 peptide (Fig. 5A); and with control peptide (Fig. 5B), respectively, in cellular lesions in Mycobacterium tuberculosis Erdman infected C3HeB/FeJ mice.
  • Ranges can he expressed herein as from “about” one particular value, and/or to "about” another particular value. When such a range is expressed, another embodiment includes from the one particular value and/or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed.
  • 45652286 “reduces,” or “reduction” refer to decreases below basal levels, e.g., as compared to a control.
  • inhibitor means to reduce or decrease in activity or expression. This can be a complete inhibition of activity or expression, or a partial inhibition. Inhibition can be compared to a control or to a standard level. Inhibition can be 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10,
  • nanoparticle generally refers to a structure of any shape having a diameter from about 1 nm up to, but not including, about 1 micron, more preferably from about 5 nm to about 500 nm, most preferably about 100 nm.
  • Nanoparticles having a spherical shape are generally referred to as “nanospheres.”
  • Nonlimiting examples of nanoparticles include virus-like particles, particles formed of high- order structures of proteins, and soft nanoparticles, e.g., micelles, colloids, liposomes, vesicles, nanodroplets nano-structured hydrogel, nanocrystals, and nanosuspension.
  • Soft nanoparticles generally dissolve or dissemble to release agents.
  • Mtb and “M. tuberculosis” are used interchangeably herein, to refer to the organism Mycobacterium tuberculosis, including all strains and variants thereof.
  • Mycobacterium sp.” refers to multiple species of the bacteria Mycobacterium.
  • Mycobacterium sp. refers to a single, but unspecified species of the bacteria Mycobacterium.
  • the term “selectively target” refers to the capability of a referenced component to bind to, target (e.g., for binding), or home to (e.g., in a subject) to the referenced component’s binding target preferentially, to a greater extent, and/or at higher affinity to the referenced component’s binding target than other components, compounds, and/or materials in a milieu of interest (e.g., in a subject).
  • the disclosed microbial pathogen-targeting peptides bind to microbial pathogens and to pathogen-infected tissues in vivo to a greater extent than to uninfected “control” cells and tissues of the same animal in vivo.
  • the disclosed microbial pathogen -targeting peptides bind to Mycobacterium bacteria to a greater extent than to cells and tissues of animals.
  • the disclosed microbial pathogen -targeting peptides target and home to Mycobacterium bacteria present in a subject to a greater extent than to cells and tissues of the subject.
  • polymer refers to a chemical entity with a plurality of repeating units generally bonded covalently. In some forms, a polymer has a molecular weight greater than 500 or 1,000, or more. Non- limiting exemplary polymers include poly-amino acids, naturally occurring, and synthetic chemical compounds.
  • pharmaceutically acceptable refers to compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit/risk ratio, in accordance with the guidelines of agencies such as the Food and Drug Administration.
  • biocompatible and “biologically compatible,” as used herein, generally refer to materials that are, along with any metabolites or degradation products thereof, generally non-toxic to the recipient, and do not cause any significant adverse effects to the recipient.
  • biocompatible materials are materials which do not elicit a significant inflammatory or immune response when administered to a patient.
  • hydrophilic refers to the property of having affinity for water.
  • hydrophilic polymers or hydrophilic polymer segments
  • hydrophilic polymer segments are polymers (or polymer segments) which are primarily soluble in aqueous solutions and/or have a tendency to absorb water.
  • hydrophilic a polymer the more hydrophilic a polymer is, the more that polymer tends to dissolve in, mix with, or be wetted by water.
  • hydrophobic refers to the property of lacking affinity for or repelling water. For example, the more hydrophobic a polymer (or polymer segment), the more that polymer (or polymer segment) tends to not dissolve in, not mix with, or not be wetted by water.
  • therapeutic agent refers to an agent that can be administered to prevent or treat a disease or disorder.
  • Therapeutic agents can be a nucleic acid, a nucleic acid analog, a small molecule, a peptidomimetic, a protein, peptide, carbohydrate or sugar, lipid, or surfactant, or a combination thereof.
  • treating and “retarding development of’ a disease, disorder, or condition occurring in an animal which has or may be predisposed to the disease, disorder and/or condition mean inhibiting the disease, disorder or condition, e.g.. impeding its progress; and relieving the disease, disorder, or condition, e.g., causing regression of the
  • Treating the disease or condition includes ameliorating at least one symptom of the particular disease or condition, even if the underlying pathophysiology is not affected, such as treating the pain of a subject by administration of an analgesic agent even though such agent does not treat the cause of the pain.
  • the animal has been diagnosed with the disease or disorder. In other forms, the animal has not yet been diagnosed as having the disease or disorder.
  • preventing and “preventing development of’ used in the context of a disease, disorder, or condition in an animal mean inhibiting the initiation or development of the disease, disorder or condition, e.g., stopping the animal from developing the disease, disorder or condition, or impeding its progress; and/or preventing the advancement or continuation of at least one symptom of the particular disease or condition, even if the underlying pathophysiology is not affected, such as preventing pain in a subject by administration of an analgesic agent, even though such agent does not treat the cause of the pain.
  • targeting moiety refers to a moiety that localizes to or away from a specific locale.
  • the moiety may be, for example, a protein, nucleic acid, nucleic acid analog, carbohydrate, or small molecule.
  • the entity may be, for example, a therapeutic compound such as a small molecule, or a diagnostic entity such as a detectable label.
  • the locale may be a tissue, a particular cell type, or a subcellular compartment.
  • terapéuticaally effective amount means an amount of the therapeutic agent that, when incorporated into and/or onto particles described herein, produces some desired effect at a reasonable benefit/risk ratio applicable to any treatment.
  • the effective amount may vary depending on such factors as the disease or condition being treated, the particular formulation being administered, the size of the subject, or the severity of the disease or condition.
  • incorporated and “encapsulated” means incorporating, formulating, or otherwise including an agent into and/or onto a composition, regardless of the manner by which the agent or other material is incorporated.
  • fragment means a portion of a polypeptide or nucleic acid molecule. This portion contains, preferably, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the entire length of the reference nucleic acid molecule or polypeptide.
  • a fragment may contain 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 nucleotides or amino acids.
  • isolated means material that is free to varying degrees from components which normally accompany it as found in its native
  • Isolate denotes a degree of separation from original source or surroundings.
  • Purify denotes a degree of separation that is higher than isolation.
  • polypeptides includes proteins and fragments thereof.
  • protein or “polypeptide” or “peptide” is meant any chain of more than two natural or unnatural amino acids, regardless of post-translational modification (e.g., glycosylation or phosphorylation), constituting all or part of a naturally occurring or non-naturally occurring polypeptide or peptide, as is described herein.
  • Polypeptides are disclosed herein as amino acid residue sequences. Those sequences are written left to right in the direction from the amino to the carboxy terminus.
  • amino acid residue sequences are denominated by either a three letter or a single letter code as indicated as follows: Alanine (Ala, A), Arginine (Arg, R), Asparagine (Asn, N), Aspartic Acid (Asp, D), Cysteine (Cys, C), Glutamine (Gin, Q), Glutamic Acid (Glu, E), Glycine (Gly, G), Histidine (His, H), Isoleucine (He, I), Leucine (Leu, L), Lysine (Lys, K), Methionine (Met, M), Phenylalanine (Phe, F), Proline (Pro, P), Serine (Ser, S), Threonine (Thr, T), Tryptophan (Trp, W), Tyrosine (Tyr, Y), and Valine (Vai, V).
  • Variant refers to a polypeptide or polynucleotide that differs from a reference polypeptide or polynucleotide but retains essential properties.
  • a typical variant of a polypeptide differs in amino acid sequence from another, reference polypeptide. Generally, differences are limited so that the sequences of the reference polypeptide and the variant are closely similar overall and, in many regions, identical.
  • a variant and reference polypeptide may differ in amino acid sequence by one or more modifications e.g., substitutions, additions, and/or deletions).
  • a substituted or inserted amino acid residue may or may not be one encoded by the genetic code.
  • a variant of a polypeptide may be naturally occurring such as an allelic variant, or it may be a variant that is not known to occur naturally.
  • Modifications and changes can be made in the structure of the polypeptides of in disclosure and still obtain a molecule having similar characteristics as the polypeptide (e.g., a conservative amino acid substitution).
  • certain amino acids can be substituted for other amino acids in a sequence without appreciable loss of activity. Because it is the interactive capacity and nature of a polypeptide that defines that polypeptide’s biological functional activity, certain amino acid sequence substitutions can be made in a polypeptide sequence and nevertheless obtain a polypeptide with like properties.
  • the hydropathic index of amino acids can be considered.
  • the importance of the hydropathic amino acid index in conferring interactive biologic function on a polypeptide is generally understood in the art. It is known that certain amino acids can be substituted for other amino acids having a similar hydropathic index or score and still result in a polypeptide with similar biological activity. Each amino acid has been assigned a hydropathic index on the basis of its hydrophobicity and charge characteristics.
  • Those indices are: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine/cysteine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (- 0.7); serine (-0.8); tryptophan (-0.9); tyrosine (-1.3); proline (-1.6); histidine (-3.2); glutamate (-3.5); glutamine (-3.5); aspartate (-3.5); asparagine (-3.5); lysine (-3.9); and arginine (-4.5).
  • the relative hydropathic character of the amino acid determines the secondary structure of the resultant polypeptide, which in turn defines the interaction of the polypeptide with other molecules, such as enzymes, substrates, receptors, antibodies, antigens, and the like. It is known in the art that an amino acid can be substituted by another amino acid having a similar hydropathic index and still obtain a functionally equivalent polypeptide. In such changes, the substitution of amino acids whose hydropathic indices are within + 2 is preferred, those within + 1 are particularly preferred, and those within + 0.5 are even more particularly preferred.
  • hydrophilicity can also be made on the basis of hydrophilicity, particularly, where the biological functional equivalent polypeptide or peptide thereby created is intended for use in immunological embodiments.
  • the following hydrophilicity values have been assigned to amino acid residues: arginine (+3.0); lysine (+3.0); aspartate (+3.0 + 1); glutamate (+3.0 + 1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); proline (-0.5 + 1); threonine (-0.4); alanine (-0.5); histidine (-0.5); cysteine (- 1.0); methionine (-1.3); valine (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5); tryptophan (-3.4).
  • an amino acid can be substituted for another having a similar hydrophilicity value and still obtain a biologically equivalent, and in particular, an immunologically equivalent polypeptide.
  • substitution of amino acids whose hydrophilicity values are within + 2 is preferred, those within + 1 are particularly preferred, and those within + 0.5 are even more particularly preferred.
  • amino acid substitutions are generally based on the relative similarity of the amino acid side-chain substituents, for example, their hydrophobicity,
  • substitutions that take various of the foregoing characteristics into consideration are well known to those of skill in the art and include (original residue: exemplary substitution): (Ala: Gly, Ser), (Arg: Lys), (Asn: Gin, His), (Asp: Glu, Cys, Ser), (Gin: Asn), (Glu: Asp), (Gly: Ala), (His: Asn, Gin), (lie: Leu, Vai), (Leu: He, Vai), (Lys: Arg), (Met: Leu, Tyr), (Ser: Thr), (Thr: Ser), (Tip: Tyr), (Tyr: Trp, Phe), and (Vai: He, Leu).
  • Embodiments of this disclosure thus contemplate functional or biological equivalents of a polypeptide as set forth above.
  • embodiments of the polypeptides can include variants having about 50%, 60%, 70%, 80%, 90%, and 95% sequence identity to the polypeptide of interest.
  • Identity is a relationship between two or more polypeptide sequences, as determined by comparing the sequences. In the art, “identity” also means the degree of sequence relatedness between polypeptide as determined by the match between strings of such sequences. “Identity” can also mean the degree of sequence relatedness of a polypeptide compared to the full-length of a reference polypeptide. “Identity” and “similarity” can be readily calculated by known methods, including, but not limited to, those described in (Computational Molecular Biology, Lesk, A. M., Ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D.
  • Preferred methods to determine identity are designed to give the largest match between the sequences tested. Methods to determine identity and similarity are codified in publicly available computer programs. The percent identity between two sequences can be determined by using analysis software (e.g., Sequence Analysis Software Package of the Genetics Computer Group, Madison Wis.) that incorporates the Needelman and Wunsch, (J. Mol. Biol., 48: 443-453, 1970) algorithm (e.g., NBLAST, and XBLAST). The default parameters are used to determine the identity for the polypeptides of the present disclosure.
  • analysis software e.g., Sequence Analysis Software Package of the Genetics Computer Group, Madison Wis.
  • Needelman and Wunsch J. Mol. Biol., 48: 443-453, 1970
  • algorithm e.g., NBLAST, and XBLAST.
  • the default parameters are used to determine the identity for the polypeptides of the present disclosure.
  • a polypeptide sequence may be identical to the reference sequence, that is be 100% identical, or it may include up to a certain integer number of amino acid alterations as compared to the reference sequence such that the % identity is less than 100%.
  • Such alterations are selected from: at least one amino acid deletion,
  • the number of amino acid alterations for a given % identity is determined by multiplying the total number of amino acids in the reference polypeptide by the numerical percent of the respective percent identity (divided by 100) and then subtracting that product from said total number of amino acids in the reference polypeptide.
  • subject is meant a mammal, including, but not limited to, a human or nonhuman mammal, such as a bovine, equine, canine, ovine, or feline.
  • the subject is preferably a mammal in need of treatment, e.g. , a subject that has been diagnosed with a disease or a predisposition thereto.
  • the mammal is any mammal, e.g., a human, a primate, a mouse, a rat, a dog, a cat, a horse, as well as livestock or animals grown for food consumption, e.g., cattle, sheep, pigs, chickens, and goats.
  • the mammal is a human.
  • peptide receptors that are selectively expressed and/or up-regulated in microbial-infected tissues include p32, NRP-1, Fn-EDB, CD206, Retinoid X receptor beta (RXR-beta), alpha-V-integrin, hyaluronan, heparin sulphate and TNC-C. Therefore,
  • 45652286 molecules such as peptides, non-peptidic polymers and other molecules that selectively target and/or bind to one or more of p32, NRP-1, Fn-EDB, CD206, Retinoid X receptor beta (RXR-beta), alpha- V-integrin, hyaluronan, heparin sulphate and TNC-C are provided.
  • Molecules including peptides and non-peptidic polymers that selectively target and bind to tissues infected with microbial pathogens in vivo, as well as to microbial pathogens such as Mycobacterium tuberculosis, have been developed, and compositions thereof are provided.
  • the infected tissue-targeting molecules are conjugated with, complexed with, or otherwise are associated with one or more active agents, such as antimicrobial agents.
  • the infected tissue-targeting molecule/active agent conjugates are incorporated into or onto a delivery vehicle.
  • the infected tissuetargeting molecules are distributed at or near the solvent-exposed surfaces of the delivery vehicle.
  • the number, type, distribution and density of infected tissue-targeting molecule(s) associated with a delivery vehicle are effective to drive homing of the vehicle and any active agents to the site of pathogenic bacteria or infected tissues in vivo.
  • infected tissue-targeting molecules are embedded within or otherwise attached to the outer surface of a hollow or porous particle, for example, a porous particle encapsulating one or more active agents for delivery selectively to a site of pathogenic bacteria in vivo.
  • the compositions are typically formulated for delivery in vivo.
  • infected tissue binding molecule means a molecule, such as a polypeptide or a non-peptidic polymer, that is capable of binding to infected tissue in vivo, and/or to pathogenic microbes, such as bacterial cells, in the absence of another molecule that targets, chaperones or otherwise mediates the binding of the molecule to the infected tissues and/or bacterial cells.
  • the infected tissue binding molecules specifically bind to one or more substances that are extruded from pathogenic microorganisms, such as bacterial cells in the region of the infected tissue.
  • compositions of infected tissue-binding peptides include one or more active agents.
  • the compositions of infected tissue-binding peptides include one or more delivery vehicles.
  • the compositions of infected tissue-binding peptides include one or more linkers or coupling agents.
  • the compositions of infected tissue-binding peptides are formulated with one or more reagents into a formulation.
  • Peptides that selectively target and bind to tissues infected with Mycobacterium sp. in vivo, as well as to cells of Mycobacterium sp., for example, cells of Mycobacterium tuberculosis, and compositions thereof are provided.
  • the peptides are conjugated with, complexed with, or otherwise are associated with one or more active agents, such as antimicrobial agents.
  • the Mycobacterium sp.-infected tissue targeting peptide/active agent conjugates are incorporated into or onto a delivery vehicle.
  • the Mycobacterium sp. -infected tissue targeting peptides are distributed at or near the solvent-exposed surfaces of the delivery vehicle.
  • the number, type, distribution and density of Mycobacterium sp.- infected tissue targeting peptide(s) associated with a delivery vehicle are effective to drive homing of the vehicle and any active agents to the site of Mycobacterium tuberculosis bacteria and/or infected tissues in vivo.
  • Mycobacterium sp. -infected tissue targeting peptides are embedded within or otherwise attached to the outer surface of a hollow or porous particle, for example, a porous particle encapsulating one or more active agents for delivery selectively to a site of Mycobacterium tuberculosis bacteria in vivo.
  • the compositions are typically formulated for delivery in vivo.
  • Mycobacterium tuberculosis binding peptide as used herein means a polypeptide that is capable of binding to a Mycobacterium tuberculosis cell in the absence of another molecule that targets, chaperones or otherwise mediates the binding of the peptide to a Mycobacterium tuberculosis cell.
  • the Mycobacterium tuberculosis binding peptides specifically bind to one or more substances that are extruded from Mycobacterium tuberculosis cells in the region associated with the Mycobacterium tuberculosis cells, such as infected tissues or organs.
  • Molecules that selectively target and bind to the tissues infected with microbial pathogens in vivo, as well as to microbial pathogens themselves have been developed, and compositions thereof are provided.
  • the molecule is a peptide that selectively targets and binds to the extracellular matrix of infected tissues, in vivo.
  • compositions are typically formulated for delivery in vivo.
  • the infected tissue binding molecules specifically bind to or near to the surface of pathogenic microbial cells, such as Mycobacterium tuberculosis cells.
  • the infected tissue binding molecules include at least one infected tissue binding region.
  • the infected tissue binding molecules are non-toxic in vivo and do not cause damage or otherwise disrupt the integrity of tissue/cells they contact, or any other structure in the body.
  • polypeptides that selectively target and bind to tissues infected with bacteria have been discovered. As described herein, these polypeptides and variants of these polypeptides can be combined with other useful materials and compositions and can be used in various methods.
  • peptides that bind to p32, NRP-1, Fn-EDB, CD206, Retinoid X receptor beta (RXR-beta), alpha- V-integrin, hyaluronan, heparin sulphate and/or TNC-C are described as infected tissue-binding peptides.
  • Peptides that selectively target and bind to infected tissue and/or microbial pathogens, such as Mycobacterium tuberculosis bacteria include polypeptides, such as polypeptides having the amino sequence of any one or more of PPRRGLIKLKTS (“PL1”; SEQ ID NOT); AGRGRLVR (“PL3”; SEQ ID NO:2); GRPARPAR (“RPARPAR”; SEQ ID NO:3); CKRDLSRRC (“IP3”; SEQ ID NO:4); and AKRGARSTA (“LinTTl”; SEQ ID NO:5); CSPGAKVRC (SEQ ID NO:6); CSPGAK (SEQ ID NO:7); CRVLRSGSC (SEQ ID NO:8); YEQDPWGVKWWY (SEQ ID NO:9); KFRKAFKRFF (SEQ ID NOTO); CGNKRTR (SEQ ID NO: 11); CRGDKGPDC (SEQ ID NO: 12); CSGRRSSKC
  • the infected tissue binding polypeptides typically include from about eight to about twelve amino acid residues, inclusive. Therefore, each amino acid residue within the polypeptide is located at a position from 1 to 12, typically annotated from the amino (NH2) to the carboxyl (COOH) terminus.
  • the disclosed polypeptides can also mediate targeting and delivery of compounds and
  • compositions coupled to, associated with, conjugated to, or even co-administered with the polypeptide are provided.
  • the PL1 polypeptide was derived from a 26 amino acid polypeptide discovered using peptide phage biopanning.
  • polypeptides including an amino acid sequence including (a) the sequence PPRRGLIKLKTS (SEQ ID NO: 1) or a variant of SEQ ID NO:1.
  • Peptides related to PPRRGLIKLKTS can be referred to as PL1 peptides.
  • some forms of PL1 peptides can include an amino acid sequence including the sequence PPRRGLIKLKTS (SEQ ID NO: 1) or a variant of the sequence PPRRGLIKLKTS (SEQ ID NO:1) with one, two, three, four, five, six, seven, eight, nine, or ten amino acid substitutions, wherein position 6 remains leucine and position 11 remains threonine.
  • the amino acid sequence can include the sequence PPRRGLIKLKTS (SEQ ID NO:1) or a variant of the sequence PPRRGLIKLKTS (SEQ ID NO:1) with one, two, three, four, five, six, seven, or eight amino acid substitutions.
  • the amino acid sequence can include the sequence PPRRGLIKLKTS (SEQ ID NO:1) or a variant of the sequence PPRRGLIKLKTS (SEQ ID NO:1) with one, two, three, four, five, or six amino acid substitutions. In some forms, the amino acid sequence can include the sequence PPRRGLIKLKTS (SEQ ID NO:1) or a variant of the sequence PPRRGLIKLKTS (SEQ ID NO:1) with one, two, three, or four amino acid substitutions.
  • the amino acid sequence can include the sequence PPRRGLIKLKTS (SEQ ID NO:1) or a variant of having at least 50% sequence identity with PPRRGLIKLKTS (SEQ ID NO: 1); or a variant having at least 58% sequence identity with PPRRGLIKLKTS (SEQ ID NO:1); or a variant having at least 66% sequence identity with PPRRGLIKLKTS (SEQ ID NO:1); or a variant having at least 75% sequence identity with PPRRGLIKLKTS (SEQ ID NO:1); or a variant having at least 83% sequence identity with PPRRGLIKLKTS (SEQ ID NO: 1); or a variant having at least 91% sequence identity with the sequence PPRRGLIKLKTS (SEQ ID NO:1).
  • Peptides related to PPRRGLIKLKTS (SEQ ID NO: 1) can be referred to as PL1 peptides.
  • Another type of disclosed peptide is based on the amino acid sequence AGRGRLVR (SEQ ID NO:2).
  • Peptides related to AGRGRLVR (SEQ ID NO:2) can be referred to as PL3 peptides. Analysis revealed that many of the amino acids can be substituted with the peptide retaining useful binding ability. Amino acid positions 3, 5, 6,
  • some forms of PL3 peptides can include an amino acid sequence including the sequence AGRGRLVR (SEQ ID NO:2) or a variant of the sequence AGRGRLVR (SEQ ID NO:2) with one, two, three, four, five, six, or seven amino acid substitutions, wherein position 3 remains arginine.
  • the amino acid sequence can include the sequence AGRGRLVR (SEQ ID NO:2) or a variant of the sequence AGRGRLVR (SEQ ID NO:2) with one, two, three, four, five, or six amino acid substitutions, wherein position 6 remains leucine and/or position eight remains arginine.
  • the amino acid sequence can include the sequence AGRGRLVR (SEQ ID NO:2) or a variant of the sequence AGRGRLVR (SEQ ID NO:2) with one, two, three, four, or five amino acid substitutions, wherein position 6 remains leucine, position eight remains arginine, and position 5 remains arginine.
  • the amino acid sequence can include the sequence AGRGRLVR (SEQ ID NO:2) or a variant of the sequence AGRGRLVR (SEQ ID NO:2) with one, two, three, or four amino acid substitutions.
  • the amino acid sequence can include the sequence AGRGRLVR (SEQ ID NO:2) or a variant of the sequence AGRGRLVR (SEQ ID NO:2) with one, two, or three amino acid substitutions. In some forms, the amino acid sequence can include the sequence AGRGRLVR (SEQ ID NO:2) or a variant of the sequence AGRGRLVR (SEQ ID NO:2) with one or two amino acid substitutions.
  • the amino acid sequence can include the sequence AGRGRLVR (SEQ ID NO:2) or a variant of the sequence AGRGRLVR (SEQ ID NO:2) having at least 25% sequence identity with AGRGRLVR (SEQ ID NO:2); or a variant having at least 37% sequence identity with AGRGRLVR (SEQ ID NO:2); or a variant having at least 50% sequence identity with AGRGRLVR (SEQ ID NO:2); or a variant having at least 62% sequence identity with the sequence AGRGRLVR (SEQ ID NO:2); or a variant of having at least 75% sequence identity with the sequence AGRGRLVR (SEQ ID NO:2); or a variant having at least 87% sequence identity with the sequence AGRGRLVR (SEQ ID NO:2).
  • Peptides related to AGRGRLVR (SEQ ID NO:2) can be referred to as PL3 peptides.
  • the amino acid sequence can include the formula X1-X2-X3- X4-X5-X6-X7-X8-X9-X10-X11-X12
  • the amino acid sequence can include the formula X13-X14-X15-X16-X17-X18-X19
  • the amino acid sequence includes the formula X20-X21- X22-X23-X24-X25-X26-X27, or (d) combinations thereof, wherein X (1 is leucine, wherein X7 is isoleucine, leucine, or valine, wherein X9 is leucine, isoleucine, or valine, wherein Xu is threonine, wherein X19 is arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine, wherein Xis is serine, alanine, glycine, asparag
  • X22 is arginine, lysine, or histidine
  • X25 is leucine, isoleucine, valine, or alanine
  • X27 is arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine
  • Xi, X2, X3, X4, X5, Xs, X10, X12, X13, X14, X15, Xi6, X17, X20, X21, X23, X24, and X26 are each, independently, any amino acid.
  • Peptides of the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12 are PL1 peptides.
  • Peptides of the formula X20-X21-X22-X23-X24-X25-X26-X27 are PL3 peptides.
  • the PL1 peptide has an amino acid sequence including the formula XXXXXL(X 7 )X(X 9 )XTX (SEQ ID NO:40), whereby each X is, independently, any amino acid; X7 is any one of isoleucine, leucine, or valine; and X9 is any one of leucine, isoleucine, or valine.
  • the peptide has an amino acid sequence including the formula XXXXX(Xis)(Xi9) (SEQ ID NO:41), whereby each X is, independently, any amino acid; Xis is any one of serine, alanine, glycine, asparagine, or threonine; and X19 is any one of arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine.
  • the PL3 peptide has an amino acid sequence including the formula XX(X22)XX(X2s)X(X27) (SEQ ID NO:42), whereby each X is, independently, any amino acid; X22 is any one of arginine, lysine, or histidine, wherein X25 is any one of leucine, isoleucine, valine, or alanine; and X27 is any one of arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine.
  • PL1 peptides can also be described in terms of an amino acid sequence including the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12, wherein X& is leucine, wherein X7 is isoleucine, leucine, or valine, wherein X 9 is leucine, isoleucine, or valine, wherein Xu is threonine, and wherein Xi, X2, X3, X4, X5, Xs, X10, and X12, are each, independently, any amino acid, i.e., as set forth in (SEQ ID NO:40).
  • the PL1 peptide has an amino acid sequence including the formula (XI)(X2)XX(X 5 )L(X 7 )X(X9)XTX (SEQ ID NO:43), whereby each X is, independently, any amino acid; Xi is any one of proline, glycine, alanine, serine, or asparagine; X2 is any one of proline, glycine, alanine, serine, or asparagine; X5 is any one of glycine, alanine, valine, leucine, or isoleucine; X7 is any one of isoleucine, leucine, or valine; and X9 is any one of leucine, isoleucine, or valine.
  • the PL1 peptide has an amino acid sequence including the formula (Xi)(X2)XX(X5)LLXLXTX (SEQ ID NO:40), whereby each X is, independently, any amino acid; Xi is any one of proline, glycine, alanine, serine, or asparagine; X2 is any one of proline, glycine, alanine, serine, or asparagine; and X5 is any one of glycine, alanine, valine, leucine, or isoleucine. In some forms, X2 can be proline.
  • the PL1 peptide has an amino acid sequence
  • 45652286 including the formula XPXXXL(X 7 )X(X 9 )XTX (SEQ ID NO:45), whereby each X is, independently, any amino acid; X7 is any one of isoleucine, leucine, or valine; and X9 is any one of leucine, isoleucine, or valine.
  • X3 can be arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine
  • X4 can be arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine
  • Xs can be alanine, lysine, histidine, arginine, glutamate, glutamine, tyrosine, or tryptophan
  • X10 can be alanine, lysine, histidine, arginine, glutamate, glutamine, tyrosine, or tryptophan
  • X12 can be serine, alanine, glycine, asparagine, threonine, glutamine, aspartate, or proline.
  • the PL1 peptide has an amino acid sequence including the formula XP(X3)(X4)XLX(Xs)X(Xio)T(Xi2) (SEQ ID NO:46), whereby each X is, independently, any amino acid; X is any one of arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine; X4 is any one of arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine; Xs is any one of alanine, lysine, histidine, arginine, glutamate, glutamine, tyrosine, or tryptophan; X10 is any one of alanine, lysine, histidine, arginine, glutamate, glutamine, tyrosine, or tryptophan; and X12 is any one of serine
  • Xi can be proline, glycine, or alanine
  • X3 can be arginine, lysine, or histidine
  • X4 can be arginine, lysine, or histidine
  • X5 can be glycine, alanine, or valine
  • Xs can be alanine, lysine, histidine, or arginine
  • X10 can be alanine, lysine, histidine, or arginine
  • X12 can be serine, alanine, glycine, asparagine, or threonine.
  • the PL1 peptide has an amino acid sequence including the formula (Xi)P(X3)(X 4 )(X 5 )LXXX(Xio)T(Xi2) (SEQ ID NO:47), whereby each X is, independently, any amino acid; Xi is any one of proline, glycine, or alanine; X3 is any one of arginine, lysine, or histidine; X4 is any one of arginine, lysine, or histidine; X5 is any one of glycine, alanine, or valine, wherein Xs can be alanine, lysine, histidine, or arginine; X10 is any one of alanine, lysine, histidine, or arginine; and X12 is any one of serine, alanine, glycine, asparagine, or threonine.
  • any amino acid substitution at X7 and X9 are conservative amino acid substitutions. In some forms, any amino acid substitutions are conservative amino acid substitutions. In some forms, the amino acid sequence can include the sequence PPRRGPL1KLKTS (SEQ ID NO:1).
  • PL3 peptides can also be described in terms of an amino acid sequence including the formula X20-X21-X22-X23-X24-X25-X26-X27, wherein X22 is arginine, lysine, or histidine, wherein X25 is leucine, isoleucine, valine, or alanine, wherein X27 is arginine, lysine,
  • the PL3 peptide has an amino acid sequence including the formula XX(X22)XX(X2s)X(X2?) (SEQ ID NO:48), whereby each X is, independently, any amino acid; X22 is any one of arginine, lysine, or histidine; X25 is any one of leucine, isoleucine, valine, or alanine; and X27 is any one of arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine.
  • X24 can be arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine.
  • the PL3 peptide has an amino acid sequence including the formula XX(X22)X(X24)(X2s)X(X27) (SEQ ID NO:49), whereby each X is, independently, any amino acid; X22 is any one of arginine, lysine, or histidine; X24 is any one of arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine; X25 is any one of leucine, isoleucine, valine, or alanine; and X27 is any one of arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine.
  • X21 can be glycine, alanine, valine, leucine, or isoleucine
  • X23 can be glycine, alanine, valine, leucine, or isoleucine and wherein X26 can be valine, leucine, isoleucine, glycine, or alanine.
  • the PL3 peptide has an amino acid sequence including the formula X(X 2 i)(X22)(X23)(X24)(X2 5 )(X26)(X27) (SEQ ID NO:50), whereby each X is, independently, any amino acid;
  • X21 is any one of glycine, alanine, valine, leucine, or isoleucine;
  • X22 is any one of arginine, lysine, or histidine;
  • X23 is any one of glycine, alanine, valine, leucine, or isoleucine X24 is any one of arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine;
  • X25 is any one of leucine, isoleucine, valine, or alanine;
  • X26 is any one of valine, leucine, isoleucine, g
  • X20 can be alanine, glycine, valine, leucine, or isoleucine.
  • X21 is any one of glycine, alanine, valine, leucine, or isoleucine
  • X22 is any one of arginine, lysine, or histidine
  • X23 is any one of glycine, alanine, valine, leucine, or isoleucine
  • X24 is any one of arginine, lysine, histidine, glutamate, glutamine, aspartate, asparagine, or alanine
  • X25 is any one of leucine, isoleucine
  • X22 can be arginine or lysine, wherein X25 can be leucine, isoleucine, or valine, wherein X27 can be arginine, lysine, or histidine.
  • X24 can be arginine, lysine, or histidine. In some forms, X24 can be arginine or lysine. In some forms, X22 can be arginine, wherein X25 can be leucine, wherein X27 can be arginine. In some forms, X24 can be arginine.
  • the PL3 peptide has an amino acid sequence including the formula (X2o)(X2i)R(X23)(X24)L(X26)R (SEQ ID NO:52), whereby X20 is any one of alanine, glycine, valine, leucine, or isoleucine X21 is any one of glycine, alanine, valine, leucine, or isoleucine; X23 is any one of glycine, alanine, valine, leucine, or isoleucine; and X26 is any one of valine, leucine, isoleucine, glycine, or alanine.
  • any amino acid substitution at X22, X25, and X27 are conservative amino acid substitutions. In some forms, any amino acid substitutions are conservative amino acid substitutions.
  • the amino acid sequence can include the sequence AGRGRLVR (SEQ ID NO:2). In some forms, the amino acid sequence can include the sequence AGRGRLVRAKLAAALE (SEQ ID NO:38).
  • the peptide can be less than 20 amino acids in length. In some forms, the peptide can be less than 15 amino acids in length. In some forms, the peptide can be 12 amino acids in length. In some forms, the peptide can include the sequence PPRRGLIKLKTSSNTKENSVVASLRP (SEQ ID NO:39). In some forms, the peptide is linear. In some forms, the peptide is cyclic. In some forms, the peptide is a modified peptide. In some forms, the peptide is a methylated peptide. In some forms, the methylated peptide can include a methylated amino acid segment. In some forms, the peptide is N- or C-methylated in at least one position.
  • the disclosed peptides preferably include the sequences of (1) one or more PL1 peptides, (2) one or more RPARPAR peptides, (3) one or more PL3 peptides, (4) one or more PL1 peptides and one or more RPARPAR peptides, (5) one or more PL1 peptides and one or more PL3 peptides, (6) one or more RPARPAR peptides and one or more PL3 peptides, or (7) one or more PL1 peptides, one or more RPARPAR peptides, and one or more PL3 peptides.
  • compositions preferably include (1) one or more PL1 peptides, (2) one or more RPARPAR peptides, (3) one or more PL3 peptides, (4) one or more PL1 peptides and one or more RPARPAR peptides, (5) one or more PL1 peptides and one or more PL3 peptides, (6) one or more RPARPAR peptides and one or more PL3 peptides, or (7) one or more PL1 peptides, one or more RPARPAR peptides, and one or more PL3 peptides.
  • any of the disclosed peptides can be composed of, for example, amino acids, amino acid analogs, peptide
  • Any component such as the components disclosed herein, can overlap, be adjacent to, and/or be upstream, downstream, or both of a peptide, such as an PL1 and/or PL3 and/or RPARPAR peptide.
  • a peptide such as an PL1 and/or PL3 and/or RPARPAR peptide.
  • examples of such components include accessory molecules, homing molecules, protease cleavage sites, etc. It is useful to have some components coupled to or associated with a peptide, such as an PL1 and/or PL3 and/or RPARPAR peptide to be downstream (C-terminal) of the peptide.
  • activatable peptide having an accessory protein or a homing peptide downstream of the peptide will be separated from the peptide when it is activated.
  • activatable peptides having an accessory molecule or a homing molecule downstream of the peptide will be separated from the peptide when it is activated. This can have some advantages such as making the peptide function more efficient or reducing the chance for extraneous effects of the eliminated component.
  • the peptides include changes, such as addition, deletion or substitution of one or more amino acids. Therefore, in some forms, the infected tissue binding molecules are variants of any one of SEQ ID NOs:l-37 that include or consist of 7, 8, 9, 10, 11 of more than 11 amino acids.
  • a variant of any one of SEQ ID NOs:l-37 is considered to have the function of selectively target and bind to infected tissue and/or microbial pathogens, such as Mycobacterium tuberculosis bacteria if the variant has at least 70%, 80%, 90%, 95%, 100%, or more than 100%, such as 105%, 110% or more of the binding affinity for infected tissue and/or microbial pathogens, such as Mycobacterium tuberculosis bacteria of the non-variant sequence from which it is derived.
  • infected tissue and/or microbial pathogens such as Mycobacterium tuberculosis bacteria if the variant has at least 70%, 80%, 90%, 95%, 100%, or more than 100%, such as 105%, 110% or more of the binding affinity for infected tissue and/or microbial pathogens, such as Mycobacterium tuberculosis bacteria of the non-variant sequence from which it is derived.
  • variants include one or more peptides having an amino acid sequence that is at least about 50%, 60%, 70%, 80%, 90%, or 99% identical to one or more of the respective amino acid sequences of SEQ ID NOs:l-37.
  • variants can include one or more peptides having an amino acid sequence that is at least about 50%, 60%, 70%, 80%, 90%, or 99% identical to one or more of the respective amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3.
  • the variants include one or more peptides having an amino acid sequence that is at least about 50%, 60%, 70%, 80%, 90%, or 99% identical to one or more of the respective amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3.
  • the variants include one or more peptides having an amino acid sequence that is at least about 50%, 60%, 70%, 80%, 90%, or 99% identical to one or more of the respective amino acid sequences of SEQ ID NO:1, SEQ ID
  • the variants include one or more peptides having an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO:1. In other forms, the variants include one or more peptides having an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO:2. In other forms, the variants include one or more peptides having an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO:3.
  • the infected tissue and/or microbial pathogen-targeting peptide includes the amino acid sequence of any one of PPRRGLIKLKTS (SEQ ID NO:1); AGRGRLVR (SEQ ID NO:2) and GRPARPAR (SEQ ID NO:3); CKRDLSRRC (“IP3”; SEQ ID NO:4); or AKRGARSTA (“LinTTl”; SEQ ID NO:5), or a functional variant thereof having at least 80% sequence identity to any one of SEQ ID NOs:l-5, whereby the peptide selectively binds to Mycobacterium bacteria.
  • the infected tissue and/or microbial pathogen- targeting peptides include one or more additional polypeptides contiguous with the peptide of any one of SEQ ID NOs:l-37, for example, having the schematic structure:
  • the infected tissue and/or microbial pathogen binding peptides include one or more additional polypeptides, the additional peptide(s) form a peptide bond with the residue at position 1, or position 2, or both position 1 and position 2.
  • the presence of the additional covalently bound polypeptide does not substantially reduce or otherwise diminish the function of selectively targeting and binding to infected tissue and/or microbial pathogens, such as Mycobacterium tuberculosis
  • a multiplicity of molecules of the infected tissue and/or microbial pathogen binding peptides are combined to form peptide multimers, such as repeating units of one or more of SEQ ID NOs: 1-37 or functional variants thereof.
  • multimers include a multiplicity of infected tissue and/or microbial pathogen binding peptides that independently have an amino acid sequence of any one of SEQ ID NOs: 1-37 or functional variants thereof.
  • the multimers include between two and one hundred copies of the infected tissue and/or microbial pathogen binding peptides.
  • the infected tissue and/or microbial pathogen-targeting peptide variants, or fusions, or multimers have a length of up to 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 400, 500, 1000 or 2000 amino acid residues.
  • an infected tissue and/or microbial pathogen-targeting peptide fusion or variant or multimer can have a length of at least 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100 or 200 residues.
  • an infected tissue and/or microbial pathogen-targeting peptide fusion or variant or multimer can have a length of 8 to 200 residues, 8 to 100 residues, 8 to 90 residues, 8 to 80 residues, 8 to 70 residues, 8 to 60 residues, 8 to 50 residues, 8 to 40 residues, 8 to 30 residues, 8 to 20 residues, 8 to 15 residues, 8 to 10 residues, 9 to 200 residues, 9 to 100 residues, 9 to 90 residues, 9 to 80 residues, 9 to 70 residues, 9 to 60 residues, 9 to 50 residues, 9 to 40 residues, 9 to 30 residues, 9 to 20 residues, 9 to 15 residues, 9 to 10 residues, 10 to 200 residues, 10 to 100 residues, 10 to 90 residues, 10 to 80 residues, 10 to 70 residues, 10 to 60 residues, 10 to 50 residues, 10 to 40 residues, 10 to 30 residues, 10 to 20 residues, or 10 to 15 residues.
  • the disclosed peptides can have a length of up to 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 400, 500, 1000 or 2000 residues. In particular embodiments, the disclosed peptides can have a length of at least 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100 or 200 residues.
  • the disclosed peptides can have a length of 7 to 200 residues, 7 to 100 residues, 7 to 90 residues, 7 to 80 residues, 7 to 70 residues, 7 to 60 residues, 7 to 50 residues, 7 to 40 residues, 7 to 30 residues, 7 to 20 residues, 7 to 15 residues, 7 to 10 residues, 8 to 200 residues, 8 to 100 residues, 8 to 90 residues, 8 to 80 residues, 8 to 70 residues, 8 to 60 residues, 8 to 50 residues, 8 to 40 residues, 8 to 30 residues, 8 to 20 residues, 8 to 15 residues, 8 to 10 residues, 9 to 200 residues, 9 to 100 residues, 9 to 90 residues, 9 to 80 residues, 9 to 70 residues, 9 to 60 residues, 9 to 50 residues, 9 to 40 residues, 9 to 30 residues, 9 to 20 residues, 9 to 15 residues, 9 to 10 residues, 10 to 200 residues, 10 to 100 residues, 9 to 90 residues, 9 to 80 residues, 9 to
  • a protein or peptide containing an PL1 and/or PL3 and/or RPARPAR peptide can have a length of up to 50, 100, 150, 200, 250, 300, 400, 500, 1000 or 2000 residues.
  • the protein or peptide portion of an PL1 and/or PL3 and/or RPARPAR composition can have a length of at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or 200 residues.
  • the protein or peptide containing an PL1 and/or PL3 and/or RPARPAR peptide can have a length of 7 to 200 residues, 7 to 100 residues, 7 to 90 residues, 7 to 80 residues, 7 to 70 residues, 7 to 60 residues, 7 to 50 residues, 7 to 40 residues, 7 to 30 residues, 7 to 20 residues, 7 to 15 residues, 7 to 10 residues, 8 to 200 residues, 8 to 100 residues, 8 to 90 residues, 8 to 80 residues, 8 to 70 residues, 8 to 60 residues, 8 to 50 residues, 8 to 40 residues, 8 to 30 residues, 8 to 20 residues, 8 to 15 residues, 8 to 10 residues, 9 to 200 residues, 9 to 100 residues, 9 to 90 residues, 9 to 80 residues, 9 to 70 residues, 9 to 60 residues, 9 to 50 residues, 9 to 40 residues, 9 to 30 residues, 9 to 20 residues, 9 to 15 residues, 9 to 10 residues, 9 to 200 residue
  • the disclosed conjugates can have a length of up to 50, 100, 150, 200, 250, 300, 400, 500, 1000 or 2000 residues. In particular embodiments, the disclosed conjugates can have a length of at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or 200 residues.
  • the disclosed conjugates can have a length of 10 to 200 residues, 10 to 100 residues, 10 to 90 residues, 10 to 80 residues, 10 to 70 residues, 10 to 60 residues, 10 to 50 residues, 10 to 40 residues, 10 to 30 residues, 10 to 20 residues, 10 to 15 residues, 15 to 200 residues, 15 to 100 residues, 15 to 90 residues, 15 to 80 residues, 15 to 70 residues, 15 to 60 residues, 15 to 50 residues, 15 to 40 residues, 15 to 30 residues, 15 to 20 residues, 20 to 200 residues, 20 to 100 residues, 20 to 90 residues, 20 to 80 residues, 20 to 70 residues, 20 to 60 residues, 20 to 50 residues, 20 to 40 residues or 20 to 30 residues.
  • the protein or peptide portion of an PL1 and/or PL3 and/or RPARPAR composition can have a length of up to 50, 100, 150, 200, 250, 300, 400, 500, 1000 or 2000 residues.
  • the protein or peptide portion of an PL1 and/or PL3 and/or RPARPAR composition can have a length of at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or 200 residues.
  • PL1 and/or PL3 and/or RPARPAR composition can have a length of 7 to 200 residues, 7 to 100 residues, 7 to 90 residues, 7 to 80 residues, 7 to 70 residues, 7 to 60 residues, 7 to 50 residues, 7 to 40 residues, 7 to 30 residues, 7 to 20 residues, 7 to 15 residues, 7 to 10 residues, 8 to 200 residues, 8 to 100 residues, 8 to 90 residues, 8 to 80 residues, 8 to 70 residues, 8 to 60 residues, 8 to 50 residues, 8 to 40 residues, 8 to 30 residues, 8 to 20 residues, 8 to 15 residues, 8 to 10 residues, 9 to 200 residues, 9 to 100 residues, 9 to 90 residues, 9 to 80 residues, 9 to 70 residues, 9 to 60 residues, 9 to 50 residues, 9 to 40 residues, 9 to 30 residues, 9 to 20 residues, 9 to 15 residues, 9 to 10 residues, 10 to 200 residues, 9 to 100 residues, 9 to 90
  • the disclosed compositions can have a length of up to 50, 100, 150, 200, 250, 300, 400, 500, 1000 or 2000 residues. In particular embodiments, the disclosed compositions can have a length of at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or 200 residues.
  • the disclosed compositions can have a length of 10 to 200 residues, 10 to 100 residues, 10 to 90 residues, 10 to 80 residues, 10 to 70 residues, 10 to 60 residues, 10 to 50 residues, 10 to 40 residues, 10 to 30 residues, 10 to 20 residues, 10 to 15 residues, 15 to 200 residues, 15 to 100 residues, 15 to 90 residues, 15 to 80 residues, 15 to 70 residues, 15 to 60 residues, 15 to 50 residues, 15 to 40 residues, 15 to 30 residues, 15 to 20 residues, 20 to 200 residues, 20 to 100 residues, 20 to 90 residues, 20 to 80 residues, 20 to 70 residues, 20 to 60 residues, 20 to 50 residues, 20 to 40 residues or 20 to 30 residues.
  • F/T/F&T and other disclosed peptides can be stabilized against proteolysis.
  • the stability and activity of peptides can be increased by protecting some of the peptide bonds with N-methylation or C-methylation.
  • Accessory peptides and homing peptides can also or similarly be stabilized against proteolysis.
  • the disclosed peptides can be made in the form of stabilized peptides and/or formulated as long-circulating forms.
  • a polyethylene glycol conjugate can be used.
  • the disclosed peptides and/or cargos can also be administered over a period of time.
  • disclosed peptides and/or cargos can be delivered with an osmotic pump. This can extend the permeability of the target cells and tissues. Modified forms of the
  • disclosed peptides can be used.
  • disclosed peptides can be methylated (which can stabilize the peptides against proteolysis). Stability against cleavage is desirable, except for bonds to be cleaved to activate a peptide. Modifications to the disclosed peptides generally should leave them functional. A peptide with a structural difference from naturally occurring forms of peptides can be considered a modified peptide.
  • amino acid and peptide analogs which can be incorporated into the disclosed infected tissue and/or microbial pathogen-targeting peptide variants, or fusions, or multimers.
  • D amino acids or amino acids which can be used.
  • the opposite stereoisomers of naturally occurring peptides are disclosed, as well as the stereo isomers of peptide analogs.
  • These amino acids can readily be incorporated into polypeptide chains by charging tRNA molecules with the amino acid of choice and engineering genetic constructs that utilize, for example, amber codons, to insert the analog amino acid into a peptide chain in a site specific way (Thorson et al., Methods in Molec. Biol.
  • infected tissue and/or microbial pathogen- targeting peptide fusion, or variant, or multimer molecules are produced that resemble peptides, but which are not connected via a natural peptide linkage.
  • peptide analogs can have more than one atom between the bond atoms, such as b-alanine, g- aminobutyric acid, and the like.
  • Amino acid analogs and peptide analogs often have enhanced or desirable properties, such as, more economical production, greater chemical stability, enhanced pharmacological properties (half-life, absorption, potency, efficacy, etc.), altered specificity (e.g., a broad-spectrum of biological activities), reduced antigenicity, and others.
  • D-amino acids can be used to generate more stable peptides, because D amino acids are not recognized by peptidases and such.
  • Systematic substitution of one or more amino acids of a consensus sequence with a D- amino acid of the same type e.g., D-lysine in place of L-lysine can be used to generate more stable peptides.
  • Cysteine residues can be used to cyclize or attach two or more peptides together. This can be beneficial to constrain peptides into particular conformations. (Rizo and Gierasch Ann. Rev. Biochem. 61:387 (1992), incorporated herein by reference).
  • Target receptors that are selectively expressed on tissues infected with microbial pathogens in vivo, as well as at the surface of microbial pathogens themselves have been identified as potential targets for the described molecules that target infected tissue and/or microbial pathogens.
  • the target receptor is selectively expressed at the extracellular matrix of infected tissues, in vivo.
  • the target receptor is specifically bound by one or more peptide having an amino acid sequence of any one or more of SEQ ID NOs:l-37.
  • peptide receptors in TB- infected pulmonary lesions include p32, NRP-1, Fn-EDB, CD206, Retinoid X receptor beta (RXR-beta), alpha- V-integrin, hyaluronan, heparin sulphate and TNC-C.
  • target receptors expressed in Mtb-infected tissues In some forms, target receptors are upregulated in cellular and necrotic lesions in response to an increase in the number of Mtb bacilli. Therefore, in some forms the amount or number of target receptors in a tissue or organ is directly proportional to the amount or number of Mtb bacilli within or around an infected tissue in vivo.
  • the target receptor is the protein known as p32, also known as HABP1 (hyaluronan-binding protein 1), gClqR (receptor for globular head domains complement Iq) or Clqbp (complement Iq-binding protein).
  • p32 protein is a doughnut-shaped trimer that has both mitochondrial and non-mitochondrial localization and functions, and has also been reported to be present at the cell surface and in the cell
  • Endogenous p32 protein is a mitochondrial protein in HeLa cells under control and stress conditions, and it has been shown that the overexpression of p32 increased mitochondrial fibrils. Therefore, in some forms, the target receptor is p32, and the described infected tissue binding molecules selectively bind p32 in vivo.
  • the target receptor is the immunoregulatory receptor Neuropilin- 1 protein known as NRP- 1.
  • NRPs Neuropilins
  • NRP1 and NRP2 Two homologous NRP isoforms are known to exist, namely NRP1 and NRP2, encoded by distinct neuropilin genes (Nrpl and Nrp2) which arose due to a gene duplication event. Both NRPs were originally discovered as neuronal adhesion molecules participating in semaphorin-mediated axonal guidance.
  • NRP1 neurotrophic factor 1
  • NRP2 nuclear factor 2
  • NRP1 nuclear factor receptor 1
  • NRP1 nuclear factor 1
  • the NRP1 protein includes a long N-terminal extracellular domain, followed by a transmembrane region and a very short cytosolic tail of 43-44 amino acids.
  • NRP-1 binds to the semaphorin group of ligands, as well as vascular endothelial growth factor (VEGF). Therefore, in some forms, the target receptor is NRP-1, and the described infected tissue binding molecules selectively bind NRP-1 in vivo.
  • VEGF vascular endothelial growth factor
  • the target receptor is Extra domain B splice variant of fibronectin (Fn-EDB).
  • Fn-EDB is an extracellular matrix protein (ECM) deposited by tumor- associated fibroblasts, and is associated with tumor growth, angiogenesis, and invasion
  • Fn-EDB includes a 91 amino acid domain inserted into fibronectin 1 at the primary transcript level, is a non-internalizing, insoluble, ECM-associated protein, and is a marker of tissue remodeling and angiogenesis. Therefore, in some forms, the target receptor is Fn- EDB, and the described infected tissue binding molecules selectively bind Fn-EDB in vivo.
  • the target receptor is Tenascin C (TNC-C).
  • TNC-C is a glycoprotein that in humans is encoded by the TNC gene. It is expressed in the
  • Tenascin-C is the founding member of the tenascin protein family. In the embryo it is made by migrating cells like the neural crest; it is also abundant in developing tendons, bone and cartilage. Therefore, in some forms, the target receptor is TNC-C, and the described infected tissue binding molecules selectively bind TNC-C.
  • compositions including infected tissue and/or microbial pathogen-targeting molecules are described.
  • the infected tissue and/or microbial pathogen-targeting molecules include one or more additional active agents, for example, to be targeted specifically to infected tissue, such as tissue infected by Mycobacterium tuberculosis bacteria.
  • the compositions include one or more active agents that is delivered specifically to infected tissue, such as tissue infected by Mycobacterium tuberculosis bacteria in vivo.
  • the infected tissue and/or microbial pathogen-targeting molecules that bind one or more of p32, NRP-1, Fn-EDB, CD206, Retinoid X receptor beta (RXR-beta), alpha- V-integrin, hyaluronan (hyaluronic acid), heparin sulphate or TNC-C include one or more additional active agents.
  • the infected tissue and/or microbial pathogen-targeting molecules include peptides having an amino acid sequence of any one or more of PPRRGLIKLKTS (“PL1”; SEQ ID NO:1); AGRGRLVR (“PL3”; SEQ ID NO:2); GRP ARP AR (“RPARPAR”; SEQ ID NO:3); CKRDLSRRC (“IP3”; SEQ ID NO:4); and AKRGARSTA (“LinTTl”; SEQ ID NO:5); CSPGAKVRC (SEQ ID NO:6); CSPGAK (SEQ ID NO:7); CRVLRSGSC (SEQ ID NO:8); YEQDPWGVKWWY (SEQ ID NO:9); KFRKAFKRFF (SEQ ID NOTO); CGNKRTR (SEQ ID NO: 11); CRGDKGPDC (SEQ ID NO: 12); CSGRRSSKC (SEQ ID NO: 13); CAQK (SEQ ID NO:14); TSK
  • compositions include one or more active agents that is delivered specifically to infected tissue, such as tissue infected by Mycobacterium tuberculosis bacteria in vivo.
  • compositions including one or more PL1 peptides and one or more PL3 peptides can be referred to as PL1-3 compositions.
  • Compositions including one or more PL1 peptides and one or more RPARPAR peptides can be referred to as PL1R compositions.
  • Compositions including one or more PL3 peptides and one or more RPARPAR peptides can be referred to as PL3R compositions.
  • Compositions including one or more PL1 peptides, one or more PL3 peptides, and one or more RPARPAR peptides can be referred to as PL13R compositions.
  • compositions including one or more PL1 peptides, one or more PL3 peptides, or both can be referred to as PL1/3 compositions.
  • Compositions including one or more PL1 peptides, one or more RPARPAR peptides, or both can be referred to as PL1/R compositions.
  • Compositions including one or more PL3 peptides, one or more RPARPAR peptides, or both can be referred to as PL3/R compositions.
  • Compositions including one or more PL1 peptides, one or more PL3 peptides, one or more RPARPAR peptides, or combinations thereof can be referred to as PL1/3/R compositions.
  • PL1 compositions Disclosed are PL1 compositions, PL1 conjugates, PL1 molecules, PL1 proteins, and PL1 peptides.
  • PL1 peptides are the basic feature of PL1 compositions, PL1 conjugates, PL1 molecules, PL1 proteins, and the PLlke.
  • PL1 compositions are any composition, conglomeration, conjugate, molecule, protein, peptide, etc. that includes an PL1 peptide.
  • PL1 conjugates are associations, whether covalent or non-covalent, of an PL1 peptide and one or more other elements, peptides, proteins, compounds, molecules, agents, compounds, etc.
  • an PL1 conjugate can include an PL1 peptide, PL1 protein, PL1 compound, PL1 molecule, etc.
  • PL1 molecules are molecules that include an PL1 peptide.
  • an PL1 molecule can include an PL1 protein, PL1 peptide, etc.
  • PL1 peptides, PL1 proteins, PL1 molecules, and PL1 conjugates are all forms of PL1 compositions.
  • PL1 compounds, PL1 peptides and PL1 proteins can be forms of PL1 molecules.
  • reference to an PL1 composition is intended to refer to PL1 compositions, PL1 molecules, PL1 proteins, PL1 peptides, and the like.
  • An PL1 component is any molecule, peptide, protein, compound, conjugate,
  • PL1 components include, for example, PL1 compositions, PL1 molecules, PL1 proteins, and PL1 peptides.
  • PL1 components can include one or more PL1 peptides.
  • PL3 compositions Disclosed are PL3 compositions, PL3 conjugates, PL3 molecules, PL3 proteins, and PL3 peptides.
  • PL3 peptides are the basic feature of PL3 compositions, PL3 conjugates, PL3 molecules, PL3 proteins, and the like.
  • PL3 compositions are any composition, conglomeration, conjugate, molecule, protein, peptide, etc. that includes an PL3 peptide.
  • PL3 conjugates are associations, whether covalent or non-covalent, of an PL3 peptide and one or more other elements, peptides, proteins, compounds, molecules, agents, compounds, etc.
  • an PL3 conjugate can include an PL3 peptide, PL3 protein, PL3 compound, PL3 molecule, etc.
  • PL3 molecules are molecules that include an PL3 peptide.
  • an PL3 molecule can include an PL3 protein, PL3 peptide, etc.
  • PL3 peptides, PL3 proteins, PL3 molecules, and PL3 conjugates are all forms of PL3 compositions.
  • PL3 compounds, PL3 peptides and PL3 proteins can be forms of PL3 molecules.
  • reference to an PL3 composition is intended to refer to PL3 compositions, PL3 molecules, PL3 proteins, PL3 peptides, and the like.
  • An PL3 component is any molecule, peptide, protein, compound, conjugate, composition, etc. that includes an PL3 peptide.
  • Examples of PL3 components include, for example, PL3 compositions, PL3 molecules, PL3 proteins, and PL3 peptides.
  • PL3 components can include one or more PL3 peptides.
  • RPARPAR compositions Disclosed are RPARPAR compositions, RPARPAR conjugates, RPARPAR molecules, RPARPAR proteins, and RPARPAR peptides.
  • RPARPAR peptides are the basic feature of RPARPAR compositions, RPARPAR conjugates, RPARPAR molecules, RPARPAR proteins, and the PL Ike.
  • RPARPAR compositions are any composition, conglomeration, conjugate, molecule, protein, peptide, etc. that includes an RPARPAR peptide.
  • RPARPAR conjugates are associations, whether covalent or non-covalent, of an RPARPAR peptide and one or more other elements, peptides, proteins, compounds, molecules, agents, compounds, etc.
  • an RPARPAR conjugate can include an RPARPAR peptide, RPARPAR protein, RPARPAR compound, RPARPAR molecule, etc.
  • RPARPAR molecules are molecules that include an RPARPAR peptide.
  • an RPARPAR molecule can include an RPARPAR protein, RPARPAR peptide, etc.
  • RPARPAR peptides, RPARPAR proteins, RPARPAR molecules, and RPARPAR conjugates are all forms of RPARPAR compositions.
  • RPARPAR compounds, RPARPAR peptides and RPARPAR proteins can be forms of RPARPAR molecules. Unless the
  • RPARPAR composition is intended to refer to RPARPAR compositions, RPARPAR molecules, RPARPAR proteins, RPARPAR peptides, and the PLlke.
  • An RPARPAR component is any molecule, peptide, protein, compound, conjugate, composition, etc. that includes an RPARPAR peptide.
  • RPARPAR components include, for example, RPARPAR compositions, RPARPAR molecules, RPARPAR proteins, and RPARPAR peptides.
  • RPARPAR components can include one or more RPARPAR peptides.
  • the PL1, and/or PL3, and/or RPARPAR peptide can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of an amino acid sequence, a protein, or a peptide that includes the PL1, and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can be covalently coupled or non- covalently associated with the PL1, and/or PL3, and/or RPARPAR peptide or an amino acid sequence, a protein, or a peptide that includes the PL1, and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can be separate from or overlapping with the PL1, and/or PL3, and/or RPARPAR peptide.
  • some accessory molecules are amino acid sequences. This can allow the amino acid sequence including the PL1, and/or PL3, and/or RPARPAR peptide to overlap the amino acid sequence that includes the accessory amino acid sequence.
  • the accessory peptide can be a separate entity that does not overlap with the PL1, and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can include a sequence in, for example, an PL1, and/or PL3, and/or RPARPAR peptide that binds to a specific receptor distinct from the receptor for the PL1 , and/or PL3, and/or RPARPAR peptide.
  • the amino acid sequence can include one or more accessory peptides.
  • the protein or peptide can include one or more accessory peptides.
  • the cargo does not include an accessory molecule.
  • the cargo can include one or more accessory molecules.
  • the cargo does not include an accessory peptide.
  • the cargo can include one or more accessory peptides.
  • the cargo can selectively home to cells and tissues having a microbial pathogen infection.
  • the cargo does not selectively home to cells and tissues having a microbial pathogen infection.
  • the cargo composition does not include an accessory molecule.
  • the cargo composition can include one or more accessory molecules.
  • the cargo composition does not include an accessory peptide.
  • the cargo composition can include one or more accessory peptides.
  • the cargo composition can selectively home to cells and tissues having a microbial pathogen infection, such as Mtb infection. In some forms, the cargo composition does not
  • the cargo composition can selectively home to cells and tissues having Mtb infection.
  • the peptide can be associated with one or more therapeutic agents.
  • a therapeutic agent can be a part of an amino acid sequence, a protein, or a peptide that includes the peptide.
  • the therapeutic agent can be covalently coupled or non-covalently associated with the peptide or an amino acid sequence, a protein, or a peptide that includes the peptide.
  • the therapeutic agent can be separate from or overlapping with the peptide.
  • some therapeutic agents are amino acid sequences. This can allow the amino acid sequence including the peptide to overlap the amino acid sequence that includes the therapeutic amino acid sequence.
  • the therapeutic agent can be a separate entity that does not overlap with the peptide.
  • the therapeutic agent can include a sequence in, for example, a peptide that binds to a specific receptor distinct from the target for the peptide.
  • the disclosed peptides home to specific cells (cells and tissues having a microbial pathogen infection) and many homing molecules home to the vasculature of the target tissue.
  • homing is referred to in some places herein as homing to the tissue associated with infection, or with the vasculature, to which the peptide or homing peptide may actually home.
  • a peptide or homing peptide with, for example, a protein, peptide, amino acid sequence, cargo, or cargo composition
  • the protein, peptide, amino acid sequence, cargo, or cargo composition can be targeted or can home to the target of the peptide or homing peptide.
  • the protein, peptide, amino acid sequence, cargo, or cargo composition or can be said to home to the target of the peptide or homing peptide.
  • reference to homing of a protein, peptide, amino acid sequence, cargo, cargo composition, etc. is intended to indicate that the protein, peptide, amino acid sequence, cargo, cargo composition, etc. includes or is associated with an appropriate peptide or homing peptide.
  • the peptide and the cargo are not covalently coupled or directly non-covalently associated with each other.
  • the cargo does not include a peptide.
  • the cargo can include one or more peptides.
  • the cargo does not include an PL1 peptide, an PL3 peptide, an RPARPAR peptide, or a homing peptide.
  • the cargo can include one or more PL1 peptides, PL3 peptides, RPARPAR peptides, or homing peptides.
  • the cargo can selectively home to cells and tissues having a microbial
  • the cargo does not selectively home to cells and tissues having a microbial pathogen infection.
  • the peptide and the cargo composition are not covalently coupled or directly non-covalently associated with each other.
  • the cargo composition does not include a peptide.
  • the cargo composition can include one or more peptides.
  • the cargo composition does not include an PL1 peptide, an PL3 peptide, an RPARPAR peptide, or a homing peptide.
  • the cargo composition can include one or more PL1 peptides, PL3 peptides, RPARPAR peptides, or homing peptides.
  • the cargo composition can selectively home to cells and tissues having a microbial pathogen infection. In some forms, the cargo composition does not selectively home to cells and tissues having microbial pathogen infection.
  • references to components as being “not covalently coupled” means that the components are not connected via covalent bonds (for example, that the PL1, and/or PL3, and/or RPARPAR peptide and the cargo are not connected via covalent bonds). That is, there is no continuous chain of covalent bonds between, for example, the PL1, and/or PL3, and/or RPARPAR peptide and the cargo.
  • references to components as being “covalently coupled” means that the components are connected via covalent bonds (for example, that the PL1 , and/or PL3, and/or RPARPAR peptide and the cargo composition are connected via covalent bonds). That is, there is a continuous chain of covalent bonds between, for example, the PL1, and/or PL3, and/or RPARPAR peptide and the cargo composition.
  • Components can be covalently coupled either directly or indirectly. Direct covalent coupling refers to the presence of a covalent bond between atoms of each of the components.
  • Indirect covalent coupling refers to the absence of a covalent bond between atoms of each of the components. That is, some other atom or atoms not belonging to either of the coupled components intervenes between atoms of the components. Both direct and indirect covalent coupling involve a continuous chain of covalent bonds.
  • Non-covalent association refers to association of components via non-covalent bonds and interactions.
  • a non-covalent association can be either direct or indirect.
  • a direct non-covalent association refers to a non-covalent bond involving atoms that are each respectively connected via a chain of covalent bonds to the components. Thus, in a direct non-covalent association, there is no other molecule intervening between the associated components.
  • An indirect non-covalent association refers to any chain of molecules and
  • references to components as not being “non-covalently associated” means that there is no direct or indirect non-covalent association between the components. That is, for example, no atom covalently coupled to an PL1 , and/or PL3, and/or RPARPAR peptide is involved in a non- covalent bond with an atom covalently coupled to a cargo.
  • an PL1, and/or PL3, and/or RPARPAR peptide and a cargo can be together in a composition where they are indirectly associated via multiple intervening non-covalent bonds while not being non-covalently associated as that term is defined herein.
  • an PL1, and/or PL3, and/or RPARPAR peptide and a cargo can be mixed together in a carrier where they are not directly non-covalently associated.
  • An PL1, and/or PL3, and/or RPARPAR peptide and a cargo that are referred to as not indirectly non-covalently associated cannot be mixed together in a continuous composition.
  • Reference to components (such as an PL1, and/or PL3, and/or RPARPAR peptide and a cargo) as not being “directly non-covalently associated” means that there is no direct non-covalent association between the components (an indirect non-covalent association may be present).
  • Reference to components (such as an PL1, and/or PL3, and/or RPARPAR peptide and a cargo) as not being “indirectly non- covalently associated” means that there is no direct or indirect non-covalent association between the components.
  • components can be non-covalently associated via multiple chains and paths including both direct and indirect non-covalent associations.
  • the presence a single direct non-covalent association makes the association a direct non-covalent association even if there are also indirect non- covalent associations present.
  • the presence of a covalent connection between components means the components are covalently coupled even if there are also non- covalent associations present. It is also understood that covalently coupled components that happened to lack any non-covalent association with each other are not considered to fall under the definition of components that are not non-covalently associated.
  • the cargo does not include a peptide.
  • the cargo can include a peptide.
  • the cargo does not include a homing peptide.
  • the cargo can include a homing peptide.
  • the cargo can selectively home to cells and tissues having microbial pathogen infection, for example, cells and tissues having Mtb infection.
  • the cargo does not selectively home to cells and tissues having microbial pathogen infection, for example, cells and tissues having Mtb infection.
  • the cargo can selectively home to cells and tissues having microbial pathogen infection, for example, cells and tissues having Mtb infection.
  • the cargo does not include an accessory molecule.
  • the cargo can include an accessory molecule.
  • the cargo does not include an accessory peptide.
  • the cargo can include an accessory peptide.
  • the cargo can selectively home to cells and tissues having Mtb infection, for example, cells and tissues having Mtb infection.
  • the PL1, and/or PL3, and/or RPARPAR peptide can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of an amino acid sequence, protein, peptide, conjugate, or composition that includes the PL1, and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can be covalently coupled or non-covalently associated with the PL1, and/or PL3, and/or RPARPAR peptide or an amino acid sequence, protein, peptide, conjugate, or composition that includes the PL I, and/or PL3, and/or RPARPAR peptide.
  • Accessory molecules can be any molecule, compound, component, etc.
  • useful accessory molecules include peptides, targeting molecules, affinity ligands, cell penetrating molecules, endosomal escape molecules, subcellular targeting molecules, nuclear targeting molecules. Different accessory molecules can have similar or different functions from each other.
  • Accessory molecules having similar functions, different functions, or both, can be associated an PL1, and/or PL3, and/or RPARPAR composition, PL1 , and/or PL3, and/or RPARPAR conjugate, PL1, and/or PL3, and/or RPARPAR molecule, PL1, and/or PL3, and/or RPARPAR protein, and/or PL1, and/or PL3, and/or RPARPAR peptide.
  • the PL I, and/or PL3, and/or RPARPAR peptide can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of an amino acid sequence, protein, peptide, conjugate, or composition that includes the PL1, and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can be covalently coupled or non-covalently associated with the PL1, and/or PL3, and/or RPARPAR peptide or an amino acid sequence, protein, peptide, conjugate, or composition that includes the PL1, and/or PL3, and/or RPARPAR peptide.
  • Accessory molecules can be
  • useful accessory molecules include peptides, targeting molecules, affinity ligands, cell penetrating molecules, endosomal escape molecules, subcellular targeting molecules, nuclear targeting molecules. Different accessory molecules can have similar or different functions from each other.
  • Accessory molecules having similar functions, different functions, or both, can be associated an PL1, and/or PL3, and/or RPARPAR composition, PL1, and/or PL3, and/or RPARPAR conjugate, PL1, and/or PL3, and/or RPARPAR molecule, PL1 , and/or PL3, and/or RPARPAR protein, and/or PL1 , and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can be separate from or overlapping with the PL1, and/or PL3, and/or RPARPAR peptide.
  • some accessory molecules are amino acid sequences. This can allow the amino acid sequence including the PL1, and/or PL3, and/or RPARPAR peptide to overlap the amino acid sequence that includes the accessory amino acid sequence.
  • the accessory molecule can be a separate entity that does not overlap with the PL1, and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can include a sequence in, for example, a peptide that binds to a specific receptor distinct from the receptor for the PL1, and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can be separate from or overlapping with the PL1, and/or PL3, and/or RPARPAR peptide.
  • some accessory molecules are amino acid sequences. This can allow the amino acid sequence including the PL1, and/or PL3, and/or RPARPAR peptide to overlap the amino acid sequence that includes the accessory amino acid sequence.
  • the accessory molecule can be a separate entity that does not overlap with the PL1, and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can include a sequence in, for example, a peptide that binds to a specific receptor distinct from the receptor for the PL1, and/or PL3, and/or RPARPAR peptide.
  • the PL1, and/or PL3, and/or RPARPAR peptide can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of an amino acid sequence, protein, peptide, conjugate, or composition that includes the PL1, and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can be
  • the PL1, and/or PL3, and/or RPARPAR conjugate can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of a conjugate or composition that includes the PL1, and/or PL3, and/or RPARPAR conjugate.
  • the accessory molecule can he covalently coupled or non-covalently associated with the PL1 , and/or PL3, and/or RPARPAR conjugate or a conjugate or composition that includes the PL1, and/or PL3, and/or RPARPAR conjugate.
  • the PL1, and/or PL3, and/or RPARPAR composition can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of a composition that includes the PL1, and/or PL3, and/or RPARPAR composition.
  • the accessory molecule can be covalently coupled or non-covalently associated with the PL1, and/or PL3, and/or RPARPAR composition or a composition that includes the PL1, and/or PL3, and/or RPARPAR composition.
  • the PL1, and/or PL3, and/or RPARPAR peptide can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of an amino acid sequence, protein, peptide, conjugate, or composition that includes the PL1, and/or PL3, and/or RPARPAR peptide.
  • the accessory molecule can be covalently coupled or non-covalently associated with the PL1, and/or PL3, and/or RPARPAR peptide or an amino acid sequence, protein, peptide, conjugate, or composition that includes the PL1, and/or PL3, and/or RPARPAR peptide.
  • the PL1, and/or PL3, and/or RPARPAR conjugate can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of a conjugate or composition that includes the PL1, and/or PL3, and/or RPARPAR conjugate.
  • the accessory molecule can be covalently coupled or non-covalently associated with the PL1, and/or PL3, and/or RPARPAR conjugate or a conjugate or composition that includes the PL1, and/or PL3, and/or RPARPAR conjugate.
  • the PL1, and/or PL3, and/or RPARPAR composition can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of a composition that includes the PL1, and/or PL3, and/or RPARPAR composition.
  • the accessory molecule can be covalently coupled or non-covalently associated with the PL1, and/or PL3, and/or RPARPAR composition or a composition that includes the PL1, and/or PL3, and/or RPARPAR composition.
  • the amino acid sequence can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of an amino acid sequence, protein, peptide, conjugate, or composition that includes the amino acid sequence.
  • the accessory molecule can be covalently coupled or non-covalently associated with the amino acid sequence or an amino acid sequence, protein, peptide, conjugate, or composition that includes the amino acid sequence.
  • the protein or peptide can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of a protein, peptide, conjugate, or composition that includes the peptide.
  • the accessory molecule can be covalently coupled or non-covalently associated with the peptide or a protein, peptide, conjugate, or composition that includes the peptide.
  • an accessory molecule can be a part of a protein, conjugate, or composition that includes the protein.
  • the accessory molecule can be covalently coupled or non-covalently associated with the protein or a protein, conjugate, or composition that includes the protein.
  • the conjugate can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of a conjugate or composition that includes the conjugate.
  • the accessory molecule can be covalently coupled or non-covalently associated with the conjugate or a conjugate or composition that includes the conjugate.
  • the composition can be associated with one or more accessory molecules.
  • an accessory molecule can be a part of a composition that includes the composition.
  • the accessory molecule can be covalently coupled or non-covalently associated with the composition or a composition that includes the composition.
  • the PL1, and/or PL3, and/or RPARPAR peptide can be associated with one or more peptides.
  • a peptide can be a part of an amino acid sequence, protein, peptide, conjugate, or composition that includes the PL1, and/or PL3, and/or RPARPAR peptide.
  • the peptide can be covalently coupled or non-covalently associated with the PL1, and/or PL3, and/or RPARPAR peptide or an amino acid sequence, protein, peptide, conjugate, or composition that includes the PL1, and/or PL3, and/or RPARPAR peptide.
  • the peptide can be separate from or overlapping with the PL1, and/or PL3, and/or RPARPAR peptide.
  • some peptides are amino acid sequences. This can allow the amino acid sequence including the PL1, and/or PL3, and/or RPARPAR peptide to overlap the amino acid sequence that includes the homing amino acid sequence.
  • the peptide can be a separate entity that does not overlap with the PL1, and/or PL3, and/or RPARPAR peptide.
  • the peptide can include a
  • the amino acid sequence of any one or more of SEQ ID NOs:l-37 can be associated with one or more peptides.
  • a peptide can be a part of an amino acid sequence, protein, peptide, conjugate, or composition that includes the amino acid sequence.
  • the peptide can be covalently coupled or non-covalently associated with the amino acid sequence or an amino acid sequence, protein, peptide, conjugate, or composition that includes the amino acid sequence.
  • the protein or peptide can be associated with one or more peptides.
  • a peptide can be a part of a protein, peptide, conjugate, or composition that includes the peptide.
  • the peptide can be covalently coupled or non-covalently associated with the peptide or a protein, peptide, conjugate, or composition that includes the peptide.
  • a peptide can be a part of a protein, conjugate, or composition that includes the protein.
  • the peptide can be covalently coupled or non-covalently associated with the protein or a protein, conjugate, or composition that includes the protein.
  • the conjugate can be associated with one or more peptides.
  • a peptide can be a part of a conjugate or composition that includes the conjugate.
  • the peptide can be covalently coupled or non-covalently associated with the conjugate or a conjugate or composition that includes the conjugate.
  • the composition can be associated with one or more peptides.
  • a peptide can be a part of a composition that includes the composition.
  • the peptide can be covalently coupled or non-covalently associated with the composition or a composition that includes the composition.
  • the term “cargo composition” refers to any cargo that can be used in conjunction with the infected tissue-targeting molecules.
  • One of skill in the art can determine what cargo can be coupled to an infected tissue-targeting molecule conjugate.
  • the infected tissue and/or microbial pathogen-targeting peptide conjugates disclosed herein can include an infected tissue-targeting molecule coupled to or associated with the cargo composition.
  • cargo compositions include, but are not limited to, an anti-microbial agent, diagnostic or labelling agents, therapeutic agents, pro-phylactic agents, anti-inflammatory agents, polypeptides, nucleic acid molecules, small molecules, nanoparticles, fluorophores, fluorescein, rhodamine, a radionuclide, indium- 111, technetium-99, carbon-11, carbon- 13, or a combination thereof.
  • cargo compositions associated with an infected tissue and/or microbial pathogen-targeting peptide in an infected tissue and/or microbial pathogen-targeting peptide in an anti-microbial agent, diagnostic or labelling agents, therapeutic agents, pro-phylactic agents, anti-inflammatory agents, polypeptides, nucleic acid molecules, small molecules, nanoparticles, fluorophores, fluorescein, rhodamine, a radionuclide, indium- 111, technetium-99, carbon-11, carbon- 13, or a combination thereof.
  • moiety is used broadly to mean a physical, chemical, or biological material that generally imparts a biologically useful function to a linked cargo composition.
  • a moiety can be any natural or nonnatural material including, without limitation, a biological material, such as a cell, phage or other virus; an organic chemical such as a small molecule; a nanoparticle, a radionuclide; a nucleic acid molecule or oligonucleotide; a polypeptide; or a peptide.
  • moieties that affect the target such as moieties with therapeutic effect, or that facilitate detection, visualization or imaging of the target, such as fluorescent molecule or radionuclides.
  • compositions of the disclosed infected tissue-targeting molecule conjugates can be combined, linked and/or coupled in any suitable manner.
  • moieties and homing molecules can be associated covalently or non-covalently, directly or indirectly, with or without a linker moiety.
  • the delivery vehicle when the infected tissue -targeting peptides are included within, i.e.. incorporated into the external surface of a delivery vehicle, the delivery vehicle includes (e.g., encapsulates or is complexed with) one or more active agent, such as a therapeutic agent.
  • the therapeutic agent is an antimicrobial agent, such as an antibiotic.
  • Exemplary active agents are selected from therapeutic agents, diagnostic agents, nutraceutical agents and prophylactic agents.
  • the active agents are selected from a protein, a nucleic acid, a carbohydrate, a polymer, a lipid, a small molecule, a cell, and a virus. Therefore, in some forms, nanoparticles or microparticles having attached thereto one or more infected tissue-targeting molecules encapsulate one or more active agents selected from proteins, nucleic acids, carbohydrates, polymers, lipids, small molecules, cells, and viruses, or combinations thereof.
  • nanoparticles or microparticles having attached thereto one or more infected tissue- targeting molecules each independently having an amino acid sequence of any one or more of SEQ ID NOs: 1-37, or functional variants thereof, encapsulate active agents that are therapeutic agents.
  • nanoparticles or microparticles having attached thereto one or more infected tissue-targeting molecules each independently having an amino acid sequence of any one or more of SEQ ID NOs: 1-37, or functional variants thereof, encapsulate active agents that are diagnostic and/or labelling agents.
  • nanoparticles or microparticles having attached thereto one or more infected tissue and/or microbial pathogen-targeting peptides each independently having an amino
  • the particles encapsulate a multiplicity of therapeutic, diagnostic and/or labelling agents.
  • causing a cargo to be covalently coupled or directly non- covalently associated refers to any action that results in a cargo that is not covalently coupled or directly non-covalently associated with the something else becoming or coming into the state of being covalently coupled or directly non-covalently associated with the something else. More clearly, "causing a cargo to be covalently coupled or directly non-covalently associated” with something else refers to any action that results in a cargo and the something else becoming or coming into the state of being covalently coupled or directly non-covalently associated. As an example, covalently coupling a cargo to another cargo constitutes "causing a cargo to be covalently coupled or directly non-covalently associated" with the other cargo.
  • a cargo that starts as a nonexistent concept and then is synthesized as part of a composition that includes the thing to which the cargo is to be coupled or directly associated constitutes "causing a cargo to be covalently coupled or directly non-covalently associated" with the thing.
  • causing a cargo composition to be covalently coupled or directly non-covalently associated refers to any action that results in a cargo composition that is not covalently coupled or directly non-covalently associated with the something else becoming or coming into the state of being covalently coupled or non- covalently associated with the something else. More clearly, “causing a cargo composition to be covalently coupled or directly non-covalently associated” with something else refers to any action that results in a cargo composition and the something else becoming or coming into the state of being covalently coupled or directly non-covalently associated.
  • covalently coupling a cargo composition to another cargo composition constitutes "causing a cargo composition to be covalently coupled or directly non- covalently associated" with the other cargo composition.
  • a cargo composition that starts as a nonexistent concept and then is synthesized as part of a composition that includes the thing to which the cargo composition is to be coupled or directly associated constitutes "causing a cargo composition to be covalently coupled or directly non-covalently associated" with the thing.
  • the cargo can be, for example, a molecule, or complex of molecules with therapeutic or diagnostic applications.
  • Therapeutic cargos that can be targeted with the disclosed peptides include but are not limited to a molecule, a complex of molecules, an anti-bacterial agent, an anti-viral agent, or a combination of these.
  • Therapeutic cargos that can be targeted with the disclosed peptides include but are not limited to a therapeutic protein, compound, or composition that is effective against microbial pathogens, such as an anti-bacterial agent, an antibody that targets microbial pathogens, a functional nucleic acid or a nucleic acid analog against one or more genes of microbial pathogens, a contrast agent, an imaging agent, a label, a labeling agent, or a combination thereof.
  • Exemplary therapeutic agents include, but are not limited to, antimicrobial agents, such as beta-lactam antibiotics (including penicillins such as ampicillin, cephalosporins selected in turn from cefuroxime, cefaclor, cephalexin, cephydroxil and cepfodoxime proxetil); tetracycline antibiotics (doxycycline and minocycline); macrolides antibiotics (azithromycin, erythromycin, rapamycin and clarithromycin); fluoroquinolones (moxifloxacin, ciprofloxacin, enrofloxacin, ofloxacin, gatifloxacin, levofloxacin) and norfloxacin and other inhibitors such as vancomycin, chloramphenicol, clindamyin, streptogramins, aminoglycosides, spectinomycin, spec tinamides, sulfonamides, trimethoprim, quinolones, diarylquinolines
  • the antibiotics are those associated clinically with treatment of an infection with a microbial pathogen.
  • the antibiotics are those associated clinically with treatment of Mtb infection, such as first- line oral drugs (with typically adult daily dose) Isoniazid (300 mg), Rifampin (600 mg), Rifabutin (300 mg), Pyrazinamide (30 mg/kg), Ethambutol (15 - 25 mg/kg); and injectable drugs (with typically adult daily dose): Streptomycin (15 mg/kg), Amikacin (15 mg/kg), Kanamycin (15 mg/kg), Capreomycin (15 mg/kg).
  • the antibiotics are those associated clinically with treatment of Mtb infection, such as Second-line oral drugs (with typically adult daily dose): Ofloxacin (400 mg twice daily ), Levofloxacin (500 mg once daily), Moxifloxacin (400 mg qd), Gatifloxacin (400 mg once daily), Ethionamide (250 mg twice daily or three times daily), Aminosalicylic acid (3 g once daily), and Cycloserine (250 mg twice daily or three times daily), bedaquiline, pretomanid, and linezolid.
  • Second-line oral drugs with typically adult daily dose
  • Ofloxacin 400 mg twice daily
  • Levofloxacin 500 mg once daily
  • Moxifloxacin 400 mg qd
  • Gatifloxacin 400 mg once daily
  • Ethionamide 250 mg twice daily or three times daily
  • Aminosalicylic acid 3 g once daily
  • Cycloserine 250 mg twice daily or three times daily
  • the antimicrobial agent is active against a wide range of bacteria in addition to Mycobacterium tuberculosis, including both gram positive and gram negative species.
  • the therapeutic agent is an antimicrobial that can kill or otherwise prevent viability of Mycobacterium tuberculosis.
  • the therapeutic agent is bedaquiline.
  • the nanoparticles or microparticles having attached thereto one or more peptides that selectively target and bind to Mycobacterium tuberculosis bacteria encapsulate vancomycin and optionally one or more therapeutic, diagnostic and/or labelling agents.
  • the antimicrobial agent is vancomycin.
  • Vancomycin is a glycopeptide antibiotic recommended intravenously as a treatment for complicated skin infections, bloodstream infections, endocarditis, bone and joint infections, and meningitis caused by methicillin-resistant bacteria. Blood levels may be measured to determine the correct dose.
  • Common adverse drug reactions (>1% of patients) associated with IV vancomycin include: local pain, which may be severe, and thrombophlebitis. Damage to the kidneys (nephrotoxicity) and to the hearing (ototoxicity) were side effects of the early impure versions of vancomycin.
  • vancomycin is combined for administered together with a second or further antimicrobial agent, such as Zosyn (piperacillin & tazobactam); Piperacillin; Timentin (Ticarcillin & clavulanate); Ticarcillin; Carbenicillin, or a 3 rd generation Cephalosporin, such as Ceftazidime.
  • a second or further antimicrobial agent such as Zosyn (piperacillin & tazobactam); Piperacillin; Timentin (Ticarcillin & clavulanate); Ticarcillin; Carbenicillin, or a 3 rd generation Cephalosporin, such as Ceftazidime.
  • Exemplary diagnostic and/or labelling agents are selected from a fluorescent label, a radioactive label, an infrared label, a coloring agent, paramagnetic molecules, fluorescent compounds, magnetic molecules, and radionuclides, X-ray imaging agents,
  • the diagnostic agents are agents that can be visualized in the body of the subject, or in a sample obtained from the subject.
  • the infected tissue-targeting molecules and/or conjugates thereof are associated with a vehicle for delivery of the peptide together with one or more active agents to infected tissues and bacteria such as Mycobacterium tuberculosis in vivo. Therefore, nanoparticles including infected tissue targeting peptides and one or more active agents are provided.
  • Exemplary delivery vehicles include substrates and particles, such as porous silicon particles, viral capsids, polymeric particles and liposomes.
  • Exemplary particles include dendrimers and beads, liposomes, viral capsids and virus-like particles, metallic particles such as beads and nano worms/nano rods, and porous silicon materials.
  • the delivery vehicles are nanoparticles or microparticles, sized for administration in vivo.
  • the delivery vehicle includes up to 5, or 10, or 15, or 20, or 25, or 30, or 35, or 40, or 45, or more than 50 copies, such as 60, 70, 80, 90, 100, 200, 300, 400, 500 or more than 500, such as 1000 or more, such as 5,000, 10,000, 15,000, 20,000, 50,000 or 100,000 copies of an infected tissue targeting molecule.
  • the delivery vehicles include between one and one hundred infected tissue-targeting molecule molecules, each independently having an amino acid sequence of any one or more of SEQ ID NOs:l-37 encapsulated within and/or associated with the delivery vehicle.
  • the delivery vehicle conjugated or complexed with the infected tissue binding molecules is a particle suitable for administration to a subject in vivo.
  • Microparticles, microspheres, and microcapsules are solid or semi-solid particles having a diameter of less than one millimeter, typically less than 100 microns, which can be formed of a variety of materials, including synthetic polymers, proteins, and polysaccharides.
  • Nanoparticles, nanospheres, and nano-capsules, referred to herein collectively as “nanoparticles”, are solid or semisolid particles having a diameter of less than one micrometer, typically less than 999 nanometers, which can be formed of a variety of materials, including synthetic polymers, proteins, and polysaccharides.
  • Nanoparticles and microparticles have been used in many different applications, primarily separations, diagnostics, and drug delivery.
  • therapeutic molecules are encapsulated within nanoparticles or microparticles or incorporated into a monolithic matrix, for subsequent release.
  • a number of different techniques are routinely used to make these nanoparticles or microparticles from synthetic polymers, natural polymers, proteins and polysaccharides, including phase separation, solvent evaporation, emulsification, and spray drying.
  • the polymers form the supporting structure of these nanospheres and microspheres, and the therapeutic agent of interest is incorporated into the polymer structure.
  • the diameter of an essentially spherical particle may refer to the physical or hydrodynamic diameter.
  • the diameter of a non-spherical particle may refer preferentially to the hydrodynamic diameter.
  • the diameter of a non-spherical particle may refer to the largest linear distance between two points on the surface of the particle.
  • Mean particle size can be measured using methods known in the art, such as dynamic light scattering.
  • “Monodisperse” and “homogeneous size distribution”, are used interchangeably herein and describe a population of nanoparticles or microparticles where all of the particles are the same or nearly the same size.
  • a monodisperse distribution refers to particle distributions in which 90% of the distribution lies within 15% of the median particle size, more preferably within 10% of the median particle size, most preferably within 5% of the median particle size.
  • Exemplary polymers used for the formation of nanospheres and microspheres that are conjugated or complexed with the infected tissue binding molecules include homopolymers and copolymers of lactic acid and glycolic acid (PLGA).
  • PLGA lactic acid and glycolic acid
  • Microspheres and nanospheres produced using polymers such as this exhibit a poor loading efficiency, however, and are often only able to incorporate a small percentage of the drug of interest into the polymer structure. Therefore, substantial quantities of microspheres and/or nanospheres often are administered to achieve a desired therapeutic effect.
  • the particles are sized for uptake within the gut following oral administration, or for uptake via the mucosa of the lung following inhalation, or for administration into the bloodstream, i.e., by intravenous injection.
  • the particle may have a diameter of between about 1,000 nm and about
  • the nanoparticle can have a diameter from 10 nm to 900 nm, from 10 nm to 800 nm, from 10 nm to 700 nm, from 10 nm to 600 nm, from 10 nm to 500 nm, from 20 nm from 500 nm, from 30 nm to 500 nm, from 40 nm to 500 nm, from 50 nm to 500 nm, from 60 nm to 400 nm, from 50 nm to 350 nm, from 50 nm to 300 nm, or from 50 nm to 200 nm.
  • the nanoparticles can have a diameter less than 400 nm, less than 300 nm, or less than 200 nm.
  • the preferred range for a nanoparticle is between 50 nm and 300 nm, or 25 nm and 250 nm, or 80 nm and 150 nm, inclusive. In particular forms a nanoparticle has a diameter of about 100 nm.
  • the particle or nanoparticle can have a zeta potential between -100 mV and +100 mV, inclusive, between -50 mV and +50 mV, inclusive, between -40 mV and +40 mV, inclusive, between -30 mV and +30 mV, inclusive, between -20 mV and +20 mV, inclusive, between -10 mV and +10mV, inclusive, or between -5mV and +5 mV, inclusive.
  • the particle or nanoparticle can have a negative zeta potential.
  • the particle can have a positive zeta potential. In some forms the particle has a substantially neutral zeta potential, i.e., the zeta potential is approximately 0 mV.
  • the particle has a zeta potential of approximately between -20 mV and +20 mV, inclusive, more preferably between -10 mV and +10 mV, inclusive. In some forms, the zeta potential is between 0 and +100, inclusive, e.g., between 0 mV and +40 mV, inclusive.
  • the delivery vehicle that is conjugated or complexed with the infected tissue-targeting molecules or conjugates thereof is a particle or substrate formed from partly or entirely of a porous silicon particle.
  • the delivery vehicle is a porous silicon material loaded with a meltable composition.
  • porous silicon material refers to a material that includes silicon, typically crystalline silicon, which has been treated by a process that has introduced a plurality of void spaces or pores into the material.
  • a “porous silicon material” includes some limited portion that is not silicon, such as carbon.
  • a “porous silicon material” can include any shape or shapes.
  • a “porous silicon material” can include porous silicon films, porous silicon layers, porous silicon particles, etc.
  • the porous silicon material includes silicon oxide or silicon dioxide. Therefore, in some forms, the delivery vehicle is a particle formed partly or completely from a “porous silicon dioxide
  • porous silicon material has been oxidized so that one or more surfaces of the porous silicon material includes silicon dioxide or silica.
  • porous silicon dioxide or “porous silica” refers to porous silicon materials or particles that have been oxidized so that the materials or particles thereof include from about 1 % up to about 99% inclusive of silicon oxide.
  • the delivery vehicle is a particle formed partly or completely from a “porous silica material” that includes some portion that is not silica, e.g., silicon.
  • the delivery vehicle is a particle formed partly or completely from a porous silicon material prepared using a sol-gel process from appropriate soluble precursors.
  • a porous silicon material prepared using a sol-gel process from appropriate soluble precursors.
  • the porous silicon oxide or porous silicon dioxide is prepared using a sol-gel process
  • the resultant porous silicon material may be referred to as a silica gel.
  • the delivery vehicle is a particle formed partly or completely from a material including silicon and optionally other elements, and which includes a plurality of void spaces or pores in the material.
  • a “porous silica material” as used herein includes structures that have an internal “core” of silicon with an outer “sheath” of silica.
  • a “porous silicon material” refers to a porous silicon-based material that contains a portion of silicon that is in its elemental form, that is, not oxidized, and a portion of silicon that has been oxidized so that one or more surfaces of the porous silicon material includes silicon dioxide.
  • a “porous silicon material” may further include some limited portion of which is not silicon, e.g., carbon.
  • the void spaces or pores within a delivery vehicle that is a particle formed partly or completely from a material including silicon are between 1 nm to 100 pm in diameter or size.
  • the dimensions of the void spaces or pores are generally tunable, i.e., the dimensions of the pores of the porous silicon material can be controlled to have a pore size dimension (e.g., diameter) of 1 nm, 5 nm, 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 70 nm, 80 nm, 100 nm, 110 nm, 120 nm, 130 nm, 140 nm, 150 nm, 200 nm, 300 nm, 400 nm, 500 nm, 1 pm, 10 pm, 50 pm, 100 pm, or any range of pore sizes that includes or is between any two of the foregoing percentages, including fractions thereof.
  • a pore size dimension e.g., diameter
  • the size of a pore within the porous silicon material is varied in diameter e.g., one side of a porous silicon material may have substantially the same size pores while the opposite side of the porous silicon material may have pores that are substantially larger, by about 25% or greater).
  • the overall porosity density of the porous silicon material is user-defined, such that pore
  • the delivery vehicle is a particle formed partly or completely from a porous silicon material having a porosity of 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or any range of porosity that includes or is between any two of the foregoing percentages, including fractions thereof.
  • the delivery vehicles formed partly or completely from a material including silicon porous silicon structures can easily withstand temperatures well in excess of what is required for wet or dry heat sterilization.
  • the delivery vehicles formed partly or completely from a material including silicon porous silicon materials readily tolerate heat sterilization without impacting their physical properties.
  • Porous silicon materials have low toxicity profiles and excellent drug delivery properties. See, e.g., PCT International Publication Nos. W02006/050221 and W02009/009563. Under biological conditions, the primary degradation product of these materials is orthosilicic acid, a non-toxic water- soluble compound naturally found in human tissues that is readily cleared through the renal system.
  • porous silicon materials for use as delivery vehicles are described in United States patent application publication number US/20200009053 Al ; US/20200397698 Al; and US/20210000744 Al, the contents of which are hereby incorporated by reference herein in their entirety.
  • delivery vehicles formed partly or completely from a porous silicon material exhibit a highly aligned nature of the pores, which results in anisotropic dissolution of an associated active agent/carrier matrix.
  • a carrier dissolves preferentially along the pore direction, guiding drug release into a zeroorder kinetic regime that result in a linear release profile. Therefore, the concentration of the free active agent is constant throughout the release period, and rapidly tapers at the end of the release period.
  • the delivery vehicle is a particle formed partly or completely from a porous silicon material that minimizes the initial burst release of an associated active agent.
  • the delivery vehicle is a particle formed partly or completely from a porous silicon material configured to deliver a constant amount of an associated active agent to a subject in vivo over a set period of time, such as one day, two days, three days, four days, five days, six days, one week, ten days, two weeks, three weeks, four weeks, one month, two months,
  • a porous silicon material has a porosity from about 15% to about 85%, inclusive, wherein the pores of the silicon material are loaded with a mixture including a thermally unstable therapeutic agent and a thermally unstable substance, wherein the thermally unstable therapeutic agent and the thermally unstable substance are not the same compound or molecule, and wherein the mixture has a lower melting point than the melting point of the thermally unstable therapeutic agent.
  • the porous silicon materials are porous silicon particles.
  • the porous silicon particles of have an average diameter or length from about 10 nm to about 100 pm.
  • the porous silicon particles of the disclosure have an average diameter or length from about 10 nm to about 100 nm. In an alternate form, the porous silicon particles have an average diameter or length from about 100 nm to about 100 pm. In a certain form, the porous silicon material has a porosity from about 50% to about 80%. In another form, the porous silicon material has a porosity of about 75%. In yet another form, the porous silicon material includes pores that have average diameters from about 2 nm to about 250 nm.
  • pSiNPs porous silicon nanoparticles
  • pSiNPs are prepared by electrochemical etch of crystalline, (100) -oriented silicon wafers (p-type, boron-doped, ⁇ 1 m.Q cm resistivity).
  • the silicon wafer is anodized in an electrolyte composed of 3:1 (v:v) of 48% aqueous HF:ethanol.
  • the etching waveform included a square wave in which a lower current density of 50 mA cm-2 is applied for 1.8 s, followed by a higher current density pulse of 400 mA cm-2 for 0.36 s.
  • the pulsed waveform is repeated for 140 cycles, and the resulting porous silicon nanostructure is lifted-off from the silicon substrate by application of a current density of 3.7 mA cm— 2 for 250 seconds in an electrolyte including 1:30 (v:v) of 48% aqueous HF:ethanol.
  • the freestanding porous silicon films are then fractured into nanoparticles of mean diameter 170 nm by ultrasonication in deionized water overnight.
  • the delivery vehicle that is conjugated or complexed with the infected tissue-targeting molecules or conjugates thereof is a particle or substrate formed from partly or entirely of a silver particle.
  • the delivery vehicle is a silver nanoparticle loaded with the protein and one or more therapeutic and/or diagnostic agents.
  • Silver nanoparticles are known in the art (see, e.g., Lingasamy et al., 2020; and Tobi et al., 2021, the contents of which are incorporated herein in their entirety).
  • the silver particles are citric AgNP.
  • Exemplary citric AgNP have an average core size of between about 5 nm and about 100 nm, inclusive such as between 10 nm and 90 nm, between 20 nm and 80 nm, or between 40 nm and 80 nm, such as 50 nm, 60 nm or 70 nm.
  • the citric AgNP has an average core size of about 62 ⁇ 20 nm.
  • Exemplary citric AgNP have an average hydrodynamic size of between about 10 nm and about 200 nm, inclusive such as between 20 nm and 160 nm, between 40 nm and 150 nm, or between 60 nm and 140 nm, such as 70 nm, 80 nm, 90 nm, 100 nm, 110 nm, 120 nm, 130 nm, or 140 nm.
  • the citric AgNP have a hydrodynamic size of 103 ⁇ 40 nm.
  • the citric AgNP have an average core size of about 62 ⁇ 20 nm, and hydrodynamic size of 103 ⁇ 40 nm.
  • AgNPs have been developed as a model platform for targeted in vitro and in vivo delivery.
  • the AgNPs have several unique features that make them useful for biodistribution studies including plasmonic enhancement emission from coupled fluorescent dyes to allow ultrasensitive imaging of single nanoparticles.
  • An exemplary citric AgNP is depicted in Figure 3C.
  • the particle includes a silver (Ag) core, covered with NeutrAvidin- PEG(5K)-thiols and Iipoic-PEG(1K)-NH2.
  • the infected tissue-targeting molecules are attached to NeutrAvidin and the payload is conjugated to the free NH2 groups of the lipoic-PEG(lK)-linker.
  • isotopically pure silver nanoparticles are synthesized and functionalized as previously described (Willmore et al., Nanoscale. 8:9096-9101 (2016)).
  • synthesis of wild type (wt) and isotopic (Agl07 and Agl09) AgNPs is carried out with biotinylated peptides being coated on NeutrAvidin (NA) on the surface of the AgNPs.
  • NA NeutrAvidin
  • transmission electron microscopy is used to image and DLS is used to assess the zeta potential, polydispersity and size of AgNPs.
  • the delivery vehicle is a viral capsid, or a virus-like particle formed from partly or entirely of a multiplicity of viral capsid proteins.
  • Preferred virus capsids are stable toward thermal denaturation at temperatures up to 80-100°C, chaotropic agents, and to extremes of pH.
  • Exemplary viral-like particles that are stable toward thermal denaturation at temperatures up to 80-100°C, chaotropic agents, and to extremes of pH include bacteriophage capsids and phage particles.
  • the delivery vehicle is a bacteriophage that infects Mycobacterium sp.
  • the delivery vehicle is a bacteriophage that infects Mycobacterium tuberculosis.
  • the delivery vehicle includes a viral-like particle (VLP), or vesicle, composed of a bacteriophage capsid protein.
  • VLP viral-like particle
  • vesicle composed of a bacteriophage capsid protein
  • the bacteriophage is an RNA or DNA phage that infects a bacteria.
  • the bacteria is Mycobacterium tuberculosis.
  • VLP virus-like particle
  • the icosahedral capsid of the RNA bacteriophage PP7 is crosslinked by disulfide bonds between coat protein dimers at its 5-fold and quasi-6-fold symmetry axes, providing enhanced stability to VLPs formed from PP7 capsid proteins.
  • the PP7 capsid is a modified PP7, for example, modified by attachment of a peptide, carbohydrate, small molecule or nucleic acid to the viral capsid. Modified PP7 VLPs are described in Zhao, et al., ACS Nano.
  • modified PP7 VLPs capable of encapsulating active agents are also described for use with the peptides.
  • the structure of the coat protein is a five-stranded P-sheet with two a-helices and a hairpin. When the capsid is assembled, the helices and hairpin face the exterior of the particle, while the 3- sheet faces the interior. d.
  • Polymeric Particles
  • the delivery vehicle is or includes one or more polymers, such as polymeric nanoparticles or microparticles.
  • Preferred polymers include biocompatible polymers.
  • the biocompatible polymer(s) is biodegradable or bioabsorbable.
  • the polymer is non-degradable.
  • the particles are a mixture of degradable and non-degradable particles.
  • the delivery vehicle is a particles that includes one or more biocompatible polymer(s) including, but not limited to, polyamino acids; cyclodextrin- containing polymers, in particular cationic cyclodextrin-containing polymers, such as those described in U.S. Patent No.
  • polymers prepared from lactones such as poly(caprolactone) (PCL); polyhydroxy acids and copolymers thereof such as poly(lactic acid) (PLA), poly(L-lactic acid) (PLLA), poly(glycolic acid) (PGA), poly(lactic acid-co- glycolic acid) (PLGA), poly(L-lactic acid-co-glycolic acid) (PLLGA), poly(D,L-lactide) (PDLA), poly(D,L-lactide-co-caprolactone), poly(D,L-lactide-co-caprolactone-co- glycolide), poly(D,L-lactide-co-PEO-co-D,L-lactide), poly(D,L-lactide-co-PPO-co-D,L- lactide), and blends thereof, polyalkyl cyanoacralate, polyurethanes, poly(valeric acid), and poly-L-glutamic acid; hydroxypropyl methacryl
  • polyesters such as polyethylene and polypropylene; polyalkylene glycols such as poly(ethylene glycol) (PEG) and polyalkylene oxides (PEG), and block copolymers thereof such as polyoxyalkylene oxide (“PLURONICS®” or block copolymers containing PEG where PEG has a molecular weight of any values within the range of 300 Daltons to 1 MDa); polyalkylene terephthalates such as poly(ethylene terephthalate); ethylene vinyl acetate polymer (EVA); polyvinyl alcohols (PVA); polyvinyl ethers; polyvinyl esters such as poly(vinyl acetate); polyvinyl halides such as poly(vinyl chloride) (PVC), polyvinylpyrrolidone; poly siloxanes; polystyrene (PS); and
  • preferred natural polymers include proteins such as albumin, collagen, gelatin and prolamines, for example, zein, and polysaccharides such as alginate. Copolymers of the above, such as random, block, or graft copolymers, or blends of the polymers listed above can also be used.
  • Functional groups on the polymer can be capped to alter the properties of the polymer and/or modify (e.g., decrease or increase) the reactivity of the functional group.
  • the carboxyl termini of carboxylic acid contain polymers, such as lactide- and glycolide-containing polymers, may optionally be capped, e.g., by esterification, and the hydroxyl termini may optionally be capped, e.g. by etherification or esterification.
  • the weight average molecular weight can vary for a given polymer but is generally from about 1000 Daltons to 1,000,000 Daltons, 1000 Daltons to 500,000 Dalton, 1000 Daltons to 250,000 Daltons, 1000 Daltons to 100,000 Daltons, 5,000 Daltons to 100,000 Daltons, 5,000 Daltons to 75,000 Daltons, 5,000 Daltons to 50,000 Daltons, or 5,000 Daltons to 25,000 Daltons.
  • the delivery vehicles are particles modified with one or more surfactants.
  • surfactants include, but are not limited to, L-a-
  • phosphatidylcholine PC
  • 1 ,2-dipalmitoylphosphatidycholine DPPC
  • oleic acid sorbitan trioleate, sorbitan mono-oleate, sorbitan monolaurate, polyoxyethylene (20) sorbitan monolaurate, polyoxyethylene (20) sorbitan monooleate, natural lecithin, oleyl polyoxyethylene (2) ether, stearyl polyoxyethylene (2) ether, lauryl polyoxyethylene (4) ether, block copolymers of oxyethylene and oxypropylene, synthetic lecithin, diethylene glycol dioleate, tetrahydrofurfuryl oleate, ethyl oleate, isopropyl myristate, glyceryl monooleate, glyceryl monostearate, glyceryl monoricinoleate, cetyl alcohol, stearyl alcohol, polyethylene glycol 400, cetyl pyri
  • PEG surface density may be controlled by varying the amount of PEG in the polymer composition or by mixing a blend of pegylated polymer component and non-pegylated polymer component.
  • the density of PEG or polyalkylene glycol on the surface of formed particles may be evaluated using several techniques.
  • the delivery vehicles are modified by the addition of one or more polymers to possess a specific ⁇ -potential.
  • the delivery vehicles are modified by the attachment of PEG and/or other polymers to the surface to possess a ⁇ -potential of between about 20 mV and about -20 mV, preferably between about 10 mV and about -10 mV, more preferably between about 2 mV and about -2 mV.
  • a ⁇ -potential of between about 20 mV and about -20 mV, preferably between about 10 mV and about -10 mV, more preferably between about 2 mV and about -2 mV.
  • the particles are lipidic particles, such as liposomes, or micelles.
  • Lipidic particles include uni-lamellar phospholipid vesicles, liposomes, or lipoprotein particles. Liposomal encapsulation may be used and the liposomes may be derivatized with various polymers (e.g., U.S. Pat. No. 5,013,556). See also Marshall, K. In: Modern Pharmaceutics Edited by G. S. Banker and C. T. Rhodes, Chapter 10, 1979.
  • Liposomes are formed from commercially available phospholipids supplied by a variety of vendors including Av anti Polar Lipids, Inc. (Birmingham, Ala.).
  • the liposome include the infected tissue binding molecules on the surface to direct the liposome to the infected tissue.
  • Liposome delivery vehicles are commercially available from multiple sources.
  • the liposome may be formed from a single lipid; however, in some embodiments, the
  • liposome is formed from a combination of more than one lipid.
  • the lipids can be neutral, anionic or cationic at physiologic pH.
  • the liposomes incorporate PEG, or PEGylated lipid derivatives. Incorporation of one or more PEGylated lipid derivatives can result in a liposome which displays polyethylene glycol chains on its surface. The resulting liposomes may possess increased stability and circulation time in vivo as compared to liposomes lacking PEG chains on their surfaces.
  • Liposomes are formed from one or more lipids, which can be neutral, anionic, or cationic at physiologic pH.
  • Suitable neutral and anionic lipids include, but are not limited to, sterols and lipids such as cholesterol, phospholipids, lysolipids, lysophospholipids, sphingolipids or pegylated lipids.
  • Neutral and anionic lipids include, but are not limited to, phosphatidylcholine (PC) (such as egg PC, soy PC), including, but limited to, 1 ,2-diacyl-glycero-3- phosphocholines; phosphatidylserine (PS), phosphatidylglycerol, phosphatidylinositol (PI); glycolipids; sphingophospholipids such as sphingomyelin and sphingoglycolipids (also known as 1-ceramidyl glucosides) such as ceramide galactopyranoside, gangliosides and cerebrosides; fatty acids, sterols, containing a carboxylic acid group for example, cholesterol; 1 ,2-diacyl-sn-glycero-3-phosphoethanolamine, including, but not limited to, 1 ,2-dioleylphosphoethanolamine (DOPE), 1 ,2-dihexadecylphospho
  • the lipids can also include various natural (e.g., tissue derived L-a-phosphatidyl: egg yolk, heart, brain, liver, soybean) and/or synthetic (e.g., saturated and unsaturated l,2-diacyl-5n-glycero-3-phosphocholines, 1-acyl- 2-acyl-.s7?-glycero-3 -phosphocholines, l,2-diheptanoyl-SN-glycero-3-phosphocholine) derivatives of the lipids.
  • the liposomes contain a phosphaditylcholine (PC) head group, and preferably sphingomyelin.
  • the liposomes contain DPPC.
  • the liposomes contain a neutral lipid, preferably 1 ,2- dioleoylphosphatidylcholine (DOPC) .
  • DOPC dioleoylphosphatidylcholine
  • the liposomes are generated from a single type of phospholipid.
  • the phospholipid has a phosphaditylcholine head group, and, most preferably is sphingomyelin.
  • the liposomes may include a sphingomyelin metabolite.
  • Sphingomyelin metabolites used to formulate the liposomes include, without limitation, ceramide, sphingosine, or sphingosine 1 -phosphate.
  • the concentration of the sphingomyelin metabolites included in the lipids used to formulate the liposomes can range from about 0.1 mol % to about 10 mol %. Preferably from about 2.0 mol % to about 5.0 mol %, and more preferably can be in a concentration of about 1.0 mol %.
  • Suitable cationic lipids in the liposomes include, but are not limited to, N-[l-(2,3- dioleoyloxy)propyl]-N,N,N-trimethyl ammonium salts, also references as TAP lipids, for example methylsulfate salt.
  • Suitable TAP lipids include, but are not limited to, DOTAP (dioleoyl-), DMTAP (dimyristoyl-), DPTAP (dipalmitoyl-), and DSTAP (distearoyl-).
  • Suitable cationic lipids in the liposomes include, but are not limited to, dimethyldioctadecyl ammonium bromide (DDAB), 1 ,2-diacyloxy-3 -trimethylammonium propanes, N-[l-(2,3-dioloyloxy)propyl]-N,N-dimethyl amine (DODAP), 1 ,2-diacyloxy-3- dimethylammonium propanes, N-[l -(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA), 1 ,2-dialkyloxy-3-dimethylammonium propanes, dioctadecylamidoglycylspermine (DOGS), 3 -[N-(N',N'-dimethylamino- ethane)carbamoyl]cholesterol (DC-Chol); 2,3-dioleoyloxy-N-(2-(
  • the cationic lipids can be l-[2-(acyloxy)ethyl]2-alkyl(alkenyl)-3-(2- hydroxyethyl)-imidazolinium chloride derivatives, for example, l-[2-(9(Z)- octadecenoyloxy)ethyl]-2-(8(Z)-heptadecenyl-3-(2-hydroxyethyl)imidazolinium chloride (DOTIM), and l-[2-(hexadecanoyloxy)ethyl]-2-pentadecyl-3-(2- hydroxyethyl)imidazolinium chloride (DPTIM).
  • DOTIM DOTIM
  • DPTIM l-[2-(hexadecanoyloxy)ethyl]-2-pentadecyl-3-(2- hydroxyethyl)imidazolinium chloride
  • the cationic lipids can be 2,3-dialkyloxypropyl quaternary ammonium compound derivatives containing a hydroxyalkyl moiety on the quaternary amine, for example, 1 ,2-dioleoyl-3-dimethyl- hydroxyethyl ammonium bromide (DORI), 1 ,2-dioleyloxypropyl-3-dimethyl- hydroxyethyl ammonium bromide (DORIE), 1 ,2-dioleyloxypropyl-3-dimetyl- hydroxypropyl ammonium bromide (DORIE-HP), 1 ,2-dioleyl-oxy-propyl-3-dimethyl- hydroxybutyl ammonium bromide (DORIE-HB), 1 ,2-dioleyloxypropyl-3 -dimethylhydroxypentyl ammonium bromide (DORIE-Hpe), 1 ,2-dimyristyloxypropyl-3-dimethylhydroxy
  • the lipids may be formed from a combination of more than one lipid, for example, a charged lipid may be combined with a lipid that is non-ionic or uncharged at
  • Non-ionic lipids include, but are not limited to, cholesterol and DOPE ( 1 ,2-dioleolylgly ceryl phosphatidylethanolamine), with cholesterol being most preferred.
  • DOPE 1 ,2-dioleolylgly ceryl phosphatidylethanolamine
  • the molar ratio of a first phospholipid, such as sphingomyelin, to second lipid can range from about 5:1 to about 1:1 or 3:1 to about 1: 1, more preferably from about 1.5:1 to about 1:1, and most preferably, the molar ratio is about 1:1.
  • compositions of infected tissue-targeting molecules or conjugates thereof include a linking chemical moiety that serves to connect, directly or indirectly, one or more moieties to the infected tissue-targeting molecules.
  • Linking chemical moieties can be any organic, inorganic, or organometallic moiety which is polyvalent, so as to provide more than two points of attachment.
  • the linking chemical moiety can be an organic molecule that contains multiple functional groups, or an organic moiety such as a substituted alkyl, unsubstituted alkyl, substituted alkylene, unsubstituted alkylene, substituted alkenyl, unsubstituted alkenyl, substituted alkynyl, unsubstituted alkynyl, substituted aryl, unsubstituted aryl, substituted cycloalkyl, unsubstituted cycloalkyl, substituted heteroalkyl, unsubstituted heteroalkyl, substituted heteroaryl, unsubstituted heteroaryl, substituted heterocyclyl, unsubstituted heterocyclyl, substituted arylalkyl, or unsubstituted arylalkyl.
  • the functional groups can be any atom or group of atoms that contains at least one atom that is neither carbon nor hydrogen, with the proviso that the groups are capable of reacting with a nucleophile or an electrophile.
  • Suitable functional groups include halogens (bromine, chlorine, and iodine); oxygen-containing functional groups such as a hydroxyls, epoxides, carbonyls, aldehydes, ester, carboxyls, and acid chlorides; nitrogen-containing functional groups such as amines and azides; and sulfur-containing groups such as thiols.
  • the functional group may also be a hydrocarbon moiety which contains one or more nonaromatic pi-bonds, such as an alkyne, alkene, or diene.
  • the linking chemical moiety can contain at least two different types of functional groups (e.g., one or more amines and one or more hydroxyls, one or more hydroxyls and one or more carboxyls, or one or more halides and one or more hydroxyls).
  • the different functional groups present on the linking chemical moiety can be independently addressed synthetically, permitting the covalent attachment of the rest of the compound and the one or more modulators in controlled stoichiometric ratios.
  • one or more moieties will be covalently joined to the linking chemical moiety via bonds, and/or
  • bonds that connect the portions of the compound to the linking chemical moiety include -C(O)NH-, -C(O)NR’-, - NR’C(O)-, -C(O)O-, -OC(O)-, -C(0)0CH2-, -SO2NR’-, -CH2R’-, -O-, -NR’H-, -NR’-, - OCONH-, -NHCOO-, -OCONR’-, -NRCOO-, -NHCONH-, -NR’CONH-, -NHCONR’-, - NR’CONR’-, -CHOH-, -CROH-, unsubstituted alkyl (such as unsubstituted C1-C12 alkyl), substituted alkyl (such as substituted C1-C12 alkyl), wherein R’ is hydrogen, halogen (F, Cl, Br, I), hydroxyl, unsubstituted al
  • the linking moiety is or includes polyethylene glycol (PEG).
  • PEGs include PEG having a molecular weight of from about 100 DA to about 20,000 Da, inclusive. In some forms, the PEG has a molecular weight of between about 1,000 Da and about 10,000 Da. In some forms, the PEG has a molecular weight of about 5,000 Da.
  • Formulations of infected tissue-targeting molecules or conjugates thereof such as compositions including delivery vehicles and active agents are also provided.
  • the compositions can be formulated for administration to a subject, for example, as a pharmaceutical formulation. Therefore, pharmaceutical formulations including a plurality of functional infected tissue-targeting molecules, each independently having an amino acid sequence of any one or more of SEQ ID NOs: 1-37, or a functional variant thereof are described.
  • Exemplary formulations include a solution, a dry powder, a tablet, micelles, colloids, nanodroplets, nano- structured hydrogel, nanocrystals, and a nanosuspension.
  • the formulation includes a determined amount of infected tissue binding molecules, in a form appropriate for a desired route of administration.
  • Exemplary formulations of infected tissue-targeting molecules including delivery vehicles and active agents include liquids and dry powders.
  • the compositions include infected tissue-targeting molecules in an amount from about 0.1% to about 50%, by weight, inclusive, from about 1% to about 40%, from about 1% to about 50%, preferably from about 1% to about 30% by weight, more
  • the infected tissue-targeting molecules to be delivered may be encapsulated within and/or associated with the surface of the particle.
  • the percent loading may be higher since the amount of is not limited by the methods of encapsulation.
  • compositions of infected tissue-targeting molecules or conjugates thereof, e.g., including nanoparticles are formulated as a liquid.
  • suitable liquid carriers include, but are not limited to, distilled water, de-ionized water, pure or ultrapure water, saline, and other physiologically acceptable aqueous solutions containing salts and/or buffers, such as phosphate buffered saline (PBS), Ringer's solution, and isotonic sodium chloride, or any other aqueous solution acceptable for administration to an animal or human.
  • PBS phosphate buffered saline
  • Ringer's solution Ringer's solution
  • isotonic sodium chloride or any other aqueous solution acceptable for administration to an animal or human.
  • Liquid formulations may include one or more suspending agents, such as cellulose derivatives, sodium alginate, polyvinylpyrrolidone, gum tragacanth, or lecithin.
  • Liquid formulations may also include one or more preservatives, such as ethyl or n-propyl p- hydroxybenzoate.
  • Formulations may be prepared using one or more pharmaceutically acceptable excipients, including diluents, preservatives, binders, lubricants, disintegrators, swelling agents, fillers, stabilizers, and combinations thereof.
  • Liquid formulations may also contain minor amounts of polymers, surfactants, or other excipients well known to those of the art.
  • minor amounts means no excipients are present that might adversely affect the delivery of the nanoparticle compositions to organs or tissues, e.g., through circulation.
  • compositions of infected tissue-targeting molecules or conjugates thereof are formulated in dry powder forms as finely divided solid formulations.
  • the dry powder components can be stored in separate containers or mixed at specific ratios and stored.
  • suitable aqueous and organic solvents are included in additional containers.
  • dry powder components, one or more solvents, and instructions on procedures to mix and prepare assembled nanostructures are included in a kit.
  • stabilized, assembled particles, nanoparticles or bulk gel thereof are dried via vacuum-drying or freeze-drying,
  • 45652286 and suitable pharmaceutical liquid carrier can be added to rehydrate and suspend the assembled nanostructures or gel compositions upon use.
  • Dry powder formulations are typically prepared by blending one or more gelators, stabilizing agents, or active agents with one or more pharmaceutically acceptable carriers.
  • Pharmaceutical carrier may include one or more dispersing agents.
  • the pharmaceutical carrier may also include one or more pH adjusters or buffers. Suitable buffers include organic salts prepared from organic acids and bases, such as sodium citrate or sodium ascorbate.
  • the pharmaceutical carrier may also include one or more salts, such as sodium chloride or potassium chloride.
  • the dry powder formulations can be suspended in the liquid formulations to form nanoparticle solutions, and administered systemically or regionally using methods known in the art for the delivery of liquid formulations.
  • compositions of infected tissue-targeting molecules or conjugates thereof, e.g., including nanoparticles are formulated for parenteral delivery, such as injection or infusion, in the form of a solution or suspension.
  • the formulation is preferably administered into the blood stream or, alternatively, directly to an organ or tissue in a subject.
  • Formulations can be prepared as aqueous compositions using techniques is known in the art.
  • such compositions can be prepared as injectable or infusible formulations, for example, solutions or suspensions; solid forms suitable for using to prepare solutions or suspensions upon the addition of a reconstitution medium prior to injection.
  • compositions of infected tissue-targeting molecules or conjugates thereof are formulated in a suitable carrier.
  • a carrier can be a solvent or dispersion medium containing, for example, water, ethanol, one or more polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), oils, such as vegetable oils (e.g., peanut oil, com oil, sesame oil, etc.), and combinations thereof.
  • compositions of infected tissue-targeting molecules or conjugates thereof are formulated to contain a preservative to prevent the growth of microorganisms.
  • Suitable preservatives include, but are not limited to, parabens, chlorobutanol, phenol, sorbic acid, and thimerosal.
  • the formulation may also contain an antioxidant to prevent degradation of the active agent(s).
  • Preservatives can be used to prevent the growth of fungi and microorganisms.
  • Suitable antifungal and antimicrobial agents include, but are not limited to, benzoic acid, butylparaben, ethyl paraben, methyl paraben, propylparaben, sodium benzoate, sodium propionate, benzalkonium chloride,
  • compositions of infected tissue-targeting molecules or conjugates thereof are formulated to be buffered to a pH, for example, pH 2, 3, 4, 5, 6, 7, 8, 9 or pH 10.
  • the formulation is typically buffered to a pH of 3-8 for parenteral administration.
  • Suitable buffers include, but are not limited to, phosphate buffers, acetate buffers, and citrate buffers.
  • compositions of infected tissue -targeting molecules or conjugates thereof are formulated to include one or more water soluble polymers.
  • Water soluble polymers are often used in formulations for parenteral administration. Suitable water-soluble polymers include, but are not limited to, polyvinylpyrrolidone, dextran, carboxymethylcellulose, and polyethylene glycol.
  • compositions of infected tissue-targeting molecules or conjugates thereof are formulated to be a sterile injectable solution.
  • Sterile injectable solutions can be prepared by incorporating the nanoparticles in the required amount in the appropriate solvent or dispersion medium with one or more of the excipients listed above, as required, followed by filtered sterilization.
  • dispersions are prepared by incorporating the various sterilized gelators, stabilizing agents, and/or active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those listed above.
  • the preferred methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
  • Formulations including compositions of infected tissue-targeting molecules or conjugates thereof may be prepared as described in standard references such as “Pharmaceutical dosage form tablets,” eds. Liberman et. al. (New York, Marcel Dekker, Inc., 1989), “Remington - The science and practice of pharmacy,” 20th ed., Lippincott Williams & Wilkins, Baltimore, MD, 2000, and “Pharmaceutical dosage forms and drug delivery systems,” 6th Edition, Ansel et al., (Media, PA: Williams and Wilkins, 1995). These references provide information on excipients, materials, equipment and process for preparing tablets and capsules and delayed release dosage forms of tablets, capsules, and granules.
  • An exemplary formulation of infected tissue-targeting molecules or conjugates thereof is a drug-loaded porous silicon particle having infected tissue-targeting molecules attached to the surface and containing one or more antibiotic agents.
  • the drug-loaded particles exhibit a temporal and linear drug release profile of an optimized formulation in vivo for an extended period of time are provided.
  • the targeted drug-loaded porous silicon particles release the entire amount of loaded drug in vivo over a period of from about two days to about 90 days, or from between about 90 days to about one year, e.g., with an overall estimated release duration in vivo of greater than 6 months.
  • the release profile showed a rapid taper at the end of the release.
  • the performance of the targeted drug delivery formulations is superior to “free” drug.
  • the targeted drug delivery formulations have reproducible syringability and exhibit a low toxicity in vivo.
  • the targeted drug delivery formulations include a porous silicon particle loaded with a meltable composition, whereby the meltable composition includes a therapeutic agent and a melting point suppression agent; whereby the meltable composition has a melting temperature; whereby the therapeutic agent has a melting temperature and a decomposition temperature; and whereby the melting temperature of the meltable composition is lower than the melting temperature and the decomposition temperature of the therapeutic agent.
  • the porous silicon nanoparticles are loaded with vancomycin.
  • vancomycin is loaded and trapped in the pSiNPs. Briefly, an aqueous dispersion of pSiNPs (1 mg ml-1 in deionized water) is added to an aqueous solution of vancomycin (0.5 mg ml-1) in a 1 :1 (v:v) ratio, and ultrasonicated for 10 min. An equal volume of aqueous calcium chloride solution (2 M) is then added to the mixture, which is agitated ( ⁇ 5 min) and ultrasonicated in an ice bath for 2 h.
  • Drug-loaded nanoparticles are purified by centrifugation and washing steps with deionized water, 70% ethanol and 100% ethanol, sequentially.
  • the infected tissue-targeting molecule including the amino acid sequence of any one of PPRRGLIKLKTS (SEQ ID NO: 1), AGRGRLVR (SEQ ID NO:2) and GRPARPAR (SEQ ID NO:3), or a functional variant thereof having at least
  • the amine-terminated nanoparticles are rinsed with ethanol and water, and then further functionalized by either MAL-PEG-SCM (maleimidepolyethylene glycol-succinimidyl carboxy methyl ester, MW 5000) or mPEG- SCM (methoxy PEG-succinimidyl carboxy methyl ester, MW 5000).
  • MAL-PEG-SCM maleimidepolyethylene glycol-succinimidyl carboxy methyl ester, MW 5000
  • mPEG- SCM methoxy PEG-succinimidyl carboxy methyl ester
  • peptide coupling is quantified and confirmed by measuring the fluorescence of a FAM label of the peptide.
  • infected tissue targeting peptides or conjugates and compositions thereof are provided.
  • the methods administer infected tissue targeting peptides or conjugates thereof associated with one or more delivery vehicles including a therapeutic antimicrobial agent to reduce or remove a pathogenic infection, such as an infection of Mycobacterium tuberculosis in a human subject.
  • infected tissue targeting peptides are useful in improving the treatment of antibiotic resistant infections. Such infections are typically treated with “last-resort” antibiotics that are typically toxic to the recipient.
  • the improvement in efficacy made possible by the described infected tissue targeting peptides reduces the unwanted side effects of such antibiotics.
  • the increase in efficacy may make it possible to overcome partial resistance of bacteria to an antibiotic.
  • infected tissue targeting peptides combine targeting with the use of nanoparticles as a delivery vector for potentially toxic antibiotics to enhance specificity of drug delivery, thereby reducing dosages and side effects.
  • the described infected tissue targeting peptides combine targeting with the use of nanoparticles as a delivery vector for potentially toxic antibiotics to enhance specificity of drug delivery, thereby reducing dosages and side effects.
  • 45652286 binding molecules have a property that may make them particularly effective in targeting infections; they not only bind to the bacteria in the infected area, but also accumulate in places, where there do not appear to be any intact bacteria. It may be that the described infected tissue binding molecules bind to a bacterial component that is released from disintegrated bacteria or shed from intact ones and that binds to tissue in the infected area. This property increases the amount of antibiotic that can be brought to the site of infection.
  • the methods treat diseases and disorders associated with microbial infections in a subject.
  • Methods of treatment for an infection by a microbial pathogen in a subject include administering to the subject a pharmaceutical formulation including
  • composition including an infected tissue-targeting molecule having an amino acid sequence of any of SEQ ID NOs:l-37, or a functional variant thereof;
  • composition (d) a pharmaceutically acceptable excipient for administration in vivo, wherein the composition is in an amount effective to kill or otherwise mitigate the microbial pathogen in the subject.
  • compositions of infected tissue targeting peptides are provided for use in treating or preventing one or more symptoms of an infection by a microbial pathogenic in a subject in need thereof, for example, including administering to the subject a composition including:
  • composition (d) a pharmaceutically acceptable excipient for administration in vivo, wherein the composition is in an amount effective to kill or otherwise mitigate the microbial pathogenic Mycobacterium sp. in the subject.
  • Methods of treating or preventing infections, such as bacterial infections in a subject are provided.
  • the methods treat or prevent one or more diseases or disorders associated with pathogenic bacterial infection.
  • An exemplary pathogenic bacterial infection is a Mycobacterium tuberculosis infection.
  • the methods administer pharmaceutical formulations including the infected tissue binding molecules to a subject in need thereof via a route selected from oral administration, intramuscular injection, intravenous injection, sub-cutaneous injection, and intra-articular injection.
  • the formulation is administered in an amount effective to reduce a microbial pathogen, such as Mycobacterium tuberculosis in the blood of the recipient to a sub- infectious amount, or amount less than or equal to between 1 and 10 cells of viable microbial pathogen per dL in blood.
  • the formulation is administered in an amount effective to reduce Mycobacterium sp., in the blood of the recipient to a sub- infectious amount, or amount less than or equal to between 1 and 10 cells of viable Mycobacterium sp. cells per dL in blood.
  • the compositions and methods typically treat or prevent infection of a subject by one or more pathogenic species of microorganisms.
  • the methods enhance the efficacy of antimicrobial agents such as antibiotics for the treatment of an infection with a pathogenic microorganism by targeting the antibiotic directly to the site of the infection following systemic administration of the composition to the subject.
  • compositions and methods of treatment thereof are useful in the context of treating and preventing infection with a microbial pathogen, and/or latent or active infectious diseases in a subject.
  • the compositions and methods treat or prevent infection of a subject by one or more pathogenic strains of Mycobacterium sp.
  • the compositions and methods treat or prevent infection of a subject by one or more pathogenic strains of Mycobacterium tuberculosis.
  • the methods treat or prevent an infection by a microbial pathogen that is a viral, fungal, bacterial or other microbial pathogen.
  • Exemplary viral pathogens include any virus including, but not limited to, a virus from any of the following viral families: Arenaviridae, Arterivirus, Astroviridae, Baculoviridae, Badnavirus, Barnaviridae, Birnaviridae, Bromoviridae, Bunyaviridae, Caliciviridae, Capillovirus, Carlavirus, Caulimovirus, Circoviridae, Closterovirus,
  • Coronaviridae e.g., Coronavirus, such as severe acute respiratory syndrome (SARS) virus
  • Corticoviridae Cystoviridae
  • Deltavirus Dianthovirus
  • Enamovirus Filoviridae (e.g., Marburg virus and Ebola virus (EBOV) (e.g., Zaire, Reston, Ivory Coast, or Sudan strain)
  • Flaviviridae (e.g., Hepatitis C virus, Dengue virus 1, Dengue virus 2, Dengue virus 3, and Dengue virus 4), Hepadnaviridae
  • Herpesviridae e.g., Human herpesvirus 1, 3, 4, 5, and 6, and Cytomegalovirus
  • Hypoviridae Iridoviridae
  • Leviviridae Lipothrixviridae
  • Microviridae Orthomyxoviridae (e.g., Influenza virus A, such as H1N1 strain, and B and C), Pa
  • Suitable viral antigens also include all or part of Dengue protein M, Dengue protein E, Dengue D INS 1 , Dengue D1NS2, and Dengue D1NS3.
  • Viral antigens may be derived from a particular strain such as a papilloma virus, a herpes virus, i.e.
  • herpes simplex 1 and 2 a hepatitis virus, for example, hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), the delta hepatitis D virus (HDV), hepatitis E virus (HEV) and hepatitis G virus (HGV), the tick-borne encephalitis viruses; parainfluenza, varicella-zoster, cytomegalovirus, Epstein- Barr, rotavirus, rhinovirus, adenovirus, coxsackieviruses, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, and lymphocytic choriomeningitis, viruses from the Orthomyxovirus family, for example, the Influenza virus A (e.g., Influenza A virus subtypes including H1N 1, H1N2, H3N2, H3N1, H5N1, H2N2, and H7N7), Influenza virus B,
  • the pathogen is a bacterial pathogen.
  • bacterial pathogens can be any pathogenic bacteria including, but not limited to bacterial strains of Actinomyces, Anabaena, Bacillus, Bacteroides, Bdellovibrio, Bordetella, Borrelia, Campylobacter, Caulobacter, Chlamydia, Chlorobium, Chromatium, Clostridium, Corynebacterium, Cytophaga, Deinococcus, Escherichia, Francisella, Halobacterium, Helicobacter, Haemophilus, Haemophilus influenza type B (HIB), Hyphomicrobium,
  • the microbial pathogen is a protozoan pathogen.
  • protozoan pathogens include but are not limited to, Cryptococcus neoformans, Histoplasma capsulation, Candida albicans, Candida tropicalis, Nocardia asteroides, Rickettsia ricketsii, Rickettsia typhi, Mycoplasma pneumoniae, Chlamydial psittaci, Chlamydial trachomatis, Plasmodium falciparum, Trypanosoma brucei, Entamoeba histolytica, Toxoplasma gondii, Trichomonas vaginalis and Schistosoma mansoni.
  • the microbial pathogen is an antibiotic-resistant microbial pathogen.
  • Antimicrobial resistance is a significant problem in some clinical strains. AMR can occur by (a) acquisition of resistance genes via horizontal gene transfer; or (b) mutations in genes already present in the genome.
  • the antibiotic-resistant strain of pathogen is a multi-drug resistant (MDR) pathogen, responsible for multi-drug resistant disease, as well as extensively drug- resistant (XDR) strains.
  • MDR multi-drug resistant
  • XDR extensively drug- resistant
  • the described infected tissue-binding molecules can effectively deliver drugs to treat an infection by an AMR pathogen that is resistant to one or more classes of antibiotics and therapeutic agents in a subject in need thereof.
  • the described compositions including the described infected tissue-binding molecules can effectively deliver a therapeutic agent in an amount effective to treat an infection by an AMR pathogen that is resistant to treatment with the same amount of the same therapeutic agent in the absence of the infected tissue homing molecules in a subject in need thereof.
  • the methods treat or prevent one or more symptoms of infection with Mycobacterium Sp.
  • the methods treat or prevent one or more symptoms of infection with one or more Mycobacterium selected from M. tuberculosis, M. leprae, M. avium, M. ulcerans, M. xenopi, M. kansasii, M. abscessus, M. chelonae, M. fortuitum, M. africanum, M. canetti, M. bovis, M. caprae, M. microti, M. pinnipedii, M. mungi, and M. orygis.
  • Mycobacterium selected from M. tuberculosis, M. leprae, M. avium, M. ulcerans, M. xenopi, M. kansasii, M. abscessus, M. chelonae, M. fortuitum, M. africanum, M. canetti, M. bovis, M. caprae, M. microti, M. pinnipedii, M. m
  • the methods treat or prevent one or more symptoms of infection with Mycobacterium tuberculosis, and/or Mycobacterium leprae in a subject in
  • the methods treat or prevent one or more symptoms of tuberculosis and/or leprosy in a subject in need thereof.
  • the methods treat or prevent treat or prevent one or more symptoms of infection with a non-tuberculosis strain of mycobacterium (“NTM”; also referred to as atypical mycobacteria, mycobacteria other than tuberculosis (MOTT), or environmental mycobacteria).
  • NTM non-tuberculosis strain of mycobacterium
  • MOTT tuberculosis
  • NTM are a large diverse cause of opportunistic lung disease that are hard to treat. NTM are common in the US where there is higher potential for more expensive treatment options. The most common type of NTM bacteria in the U.S. is Mycobacterium avium complex (MAC). The great majority of NTM lung disease in the U.S. is caused by MAC.
  • MAC Mycobacterium avium complex
  • the methods treat or prevent one or more symptoms of Mycobacterium avium complex (MAC) in a subject in need thereof.
  • MAC Mycobacterium avium complex
  • NTM lung disease Two of the other more common NTM species that infect the lungs are M. abscessus and M. kansasii.
  • Each type of NTM affects the body differently. The severity of disease, how it is treated, and the likelihood of recovery can vary widely from person to person. Some of the factors that impact the course of someone’s NTM lung disease are the virulence of the organism, the amount of exposure they’ve gotten, and their overall health. Therefore, in some forms the methods treat or prevent one or more symptoms of infection with M. abscessus and/or M. kansasii. in a subject in need thereof.
  • Mycobacterium tuberculosis(Mtb) bacteria usually attack the lungs, but TB bacteria can attack any part of the body such as the kidney, spine, and brain. Not everyone infected with TB bacteria becomes sick.
  • the methods enhance the efficacy of antimicrobial agents such as antibiotics for the treatment of an infection with a pathogenic Mycobacterium tuberculosis by targeting the antibiotic directly to the site of Mycobacterium following systemic administration of the composition to the subject.
  • compositions can also be used for treatment and/or prevention of other diseases and disorders including those associated with bacterial infection, such as secondary infections.
  • the methods treat and/or prevent a secondary infection by one or more of a virus, a bacteria, a fungus or a protozoan.
  • the subject to be treated is a human. All the methods described can include the step of identifying and selecting a subject in need of treatment, or a subject who would benefit from administration with the described compositions.
  • Secondary TB is the reactivation of a dormant infection with Mtb, usually but not always in a person with a weakened immune system. i. M. tuberculosis infection
  • the methods treat or prevent an infection of Mycobacterium tuberculosis (Mtb) in a subject in need thereof.
  • Mcb Mycobacterium tuberculosis
  • Mtb also known as Koch's bacillus, is a species of pathogenic bacteria in the family Mycobacteriaceae and the causative agent of tuberculosis.
  • M. tuberculosis has an unusual, waxy coating on its cell surface primarily due to the presence of mycolic acid.
  • the physiology of Mtb is highly aerobic and requires high levels of oxygen. Primarily a pathogen of the mammalian respiratory system, it infects the lungs.
  • the most frequently used diagnostic methods for tuberculosis are the tuberculin skin test, acid-fast stain, culture, and polymerase chain reaction. Mtb requires oxygen to grow, and is nonmotile.
  • Mtb divides every 18-24 hours, which is extremely slow compared with other bacteria, which tend to have division times measured in minutes (Escherichia coli can divide roughly every 20 minutes). Mtb is a small bacillus that can withstand weak disinfectants and can survive in a dry state for weeks. Its unusual cell wall is rich in lipids such as mycolic acid and cord factor glycolipid, is likely responsible for its resistance to desiccation and is a key virulence factor.
  • Mtb is part of a genetically related complex group of Mycobacterium species, known as Mycobacterium tuberculosis complex, including: M. tuberculosis sensu stricto; M. africanum M. canetti; M. bovis M. caprae; M. microti; M. pinnipedii; M. mungi; and M. orygis.
  • the methods treat or prevent an infection by Mycobacterium tuberculosis complex in a subject. Therefore, the methods treat and prevent infection by any one or more of M. tuberculosis sensu stricto; M. africanum; M. canetti; M. bovis; M. caprae; M. microti; M. pinnipedii; M. mungi; and M. orygis in a subject.
  • the pathogenic Mtb is an antibiotic-resistant strain of Mtb.
  • Antimicrobial resistance AMR is a significant problem in some clinical strains. AMR in M. tuberculosis typically occurs by mutations in genes already present in the genome.
  • Mtb is a multi-drug resistant Mtb (MDR-Mtb), responsible for multi-drug resistant disease (MDR-TB), as well as extensively drug -resistant (XDR-) TB.
  • MDR-Mtb multi-drug resistant Mtb
  • XDR- extensively drug -resistant
  • the pathogenic M. tuberculosis is resistant to one or more types of antibiotics.
  • the pathogenic M. tuberculosis is a multi-drug resistant (MDR) strain, that is resistant to multiple types of antibiotics, or an extensively drug resistant (XDR) strain, that is resistant to all types of antibiotics.
  • MDR multi-drug resistant
  • XDR extensively drug resistant
  • AMR Mtb that are resistant to many classes of antibiotics and therapeutic agents are problematic during infection as it can be difficult to treat. iii. Symptoms of Tuberculosis
  • the methods treat or prevent one or more symptoms of Tuberculosis (TB).
  • TB Tuberculosis
  • the methods treat or prevent one or more symptoms of pulmonary TB, CNS TB, pericardial TB, pleural TB, Lymphadenitis, Abdominal TB, skeletal TB, genitourinary TB or miliary TB in a subject in need thereof.
  • Tuberculosis can develop through progression of recently acquired infection (primary disease), reactivation of latent infection, or exogenous reinfection. In immunocompetent individuals, approximately 3-10% of those with tuberculous infection
  • the methods treat or prevent active TB in a subject in need thereof.
  • the methods treat or prevent latent TB in a subject in need thereof.
  • tuberculosis occurs as pulmonary disease with 17% occurring at an extrapulmonary site, only. However, as much as 70% of HIV- 1 infected patients will have evidence of extrapulmonary disease or mycobacteremia once the CD4 count is below 100 cells pl/ml. Co-infected persons are more likely to present atypically, potentially delaying the diagnosis of tuberculosis. iv. Pulmonary Tuberculosis
  • the methods treat or prevent one or more symptoms of pulmonary TB in a subject in need thereof.
  • Symptoms of active pulmonary TB may occur during the primary infection (and they may be relatively mild in an otherwise healthy person) or a secondary reactivation of MTB infection. These typically include: Chest pain, Wheezing or difficulty breathing, Cough (usually with mucus, sometimes with blood), Fever, sometimes with night sweats, Fatigue, and unintentional weight loss. Therefore, in some forms, the methods treat or prevent one or more symptom including Chest pain, Wheezing or difficulty breathing, Cough (usually with mucus, sometimes with blood), Fever, sometimes with night sweats, Fatigue, and unintentional weight loss in a subject having pulmonary TB.
  • CNS Central Nervous System
  • Tuberculosis of the central nervous system can present as tuberculous meningitis, tuberculomas, or tuberculous spinal meningitis.
  • the clinical spectrum of tuberculous meningitis ranges from chronic headache and subtle mental status changes to sudden, severe meningitis progressing to coma.
  • a prodrome of malaise, intermittent headache, and low grade fever can be followed by protracted headache, vomiting, confusion, meningismus, and focal neurologic signs within 2 to 3 weeks.
  • If untreated stupor, coma, seizures, and hemiparesis and death can occur within five to eight weeks after the onset of illness. Fever is not always present, and the peripheral white blood cell count is usually normal.
  • Patients may have mild anemia or hyponatremia due to inappropriate antidiuretic hormone secretion.
  • the methods treat or prevent one or more symptom including malaise, intermittent headache, fever, protracted headache, vomiting, confusion, meningismus, stupor, coma, seizures, and hemiparesis in a subject having CNS TB.
  • the methods treat or prevent one or more symptom of TB meningitis.
  • Tuberculomas are space-occupying lesions in the brain. They are usually multiple but can be single. Patients may present with seizures or other focal neurologic symptoms without evidence of systemic illness or meningeal inflammation. The meninges can become involved with encasement of the spinal cord by a gelatinous or fibrous exudates in advanced cases. Patients may have bladder or rectal sphincter weakness, hypesthesia, anesthesia, paresthesias in the distribution of a nerve root, or paralysis and pain resulting from nerve root or cord compression.
  • the methods treat or prevent one or more symptom including bladder or rectal sphincter weakness, hypesthesia, anesthesia, paresthesias in the distribution of a nerve root, or paralysis and pain resulting from nerve root or cord compression in a subject having TB meningitis.
  • the methods treat or prevent one or more symptoms of Tuberculous pleurisy in a subject in need thereof.
  • Tuberculous pleurisy can occur within weeks to months after primary infection (early postprimary pleurisy), complicate chronic pulmonary tuberculosis, or develop concurrently in 10%-30% of cases with military tuberculosis.
  • Early postprimary pleurisy usually affects adolescents and young adults. The effusion can resolve within several months in as many as 90% of cases; however, without treatment, 65% will relapse with chronic organ tuberculosis within 5 years.
  • tuberculous pleurisy may be easily mistakenly attributed to underlying comorbidities.
  • the clinical course of tuberculous pleurisy may be low grade and subtle or abrupt and severe and can be confused with acute bacterial pneumonia.
  • Patients usually have cough, pleuritic chest pain, and occasionally high fever. Night sweats, chills, weakness, dyspnea, and weight loss can also occur.
  • the effusion is usually minimal to moderate in volume and almost always unilateral (unless military tuberculosis exists concurrently).
  • the methods treat or prevent one or more symptom including cough, pleuritic chest pain, and occasionally high fever. Night sweats, chills, weakness, dyspnea, and weight loss in a subject infected with Mtb. vii. Lymphadenitis
  • the methods treat or prevent one or more symptoms of Lymphadenitis in a subject in need thereof.
  • Lymphadenitis is the most common form of extrapulmonary tuberculosis. In HIV-negative persons, it is usually unilateral and located in the cervical or supraclavicular area. The most common site is the upper border of the sternocleidomastoid muscle. Patients usually present with a painless, red, firm mass without systemic symptoms. It is most often seen in young adult females. Children often have an ongoing primary infection, but other age groups seldom have concurrent extranodal tuberculosis. Mediastinal adenopathy is often seen in children with primary infection, but it is uncommon in young adults and elderly persons.
  • Differential diagnosis of mediastinal adenopathy includes histoplasmosis, lymphoma, and cardinoma. Less commonly, tuberculosis can also cause fibrosing mediastinitis, and patients can present with dyspnea on exertion due to compression of pulmonary veins and arteries or superior vena cava syndrome. In individuals with AIDS, peripheral lymph node tuberculosis is always multifocal and associated with systemic symptoms, such as fever and weight loss. Mediastinal lymphadenopathy is frequent, and CT scan reveals multiple coalescing mediastinal masses with low-density centers, peripheral contrast enhancement, and no calcification. Abdominal lymphadenopathy in the intra- abdominal cavity is also common in AIDS patients.
  • Lymph nodes can obstruct the biliary tract, ureters, or bowel. Abscesses in the liver, spleen, pancreas, or kidney can exist concurrently. Therefore, in some forms the methods treat or prevent obstruction of the biliary tract, ureters, or bowel, and/or formation of abscesses in the liver, spleen, pancreas, or kidney in a subject with an Mtb infection, optionally also having HIV/AIDS. viii. Pericardial Tuberculosis
  • the methods treat or prevent one or more symptoms of Pericardial tuberculosis in a subject in need thereof.
  • Pericardial tuberculosis is usually caused by extension from a contiguous focus of infection, such as mediastinal or hilar nodes, the lung, spine, or sternum. Dissemination to the pericardium can occur with military tuberculosis. The onset may be abrupt or insidious. Patients may present with dyspnea, orthopnea, dull retrosternal pain, a pericardial friction rub, or symptoms and signs of cardiac tamponade. Fever, weight loss
  • the methods treat or prevent one or more of dyspnea, orthopnea, dull retrosternal pain, a pericardial friction rub, or symptoms and signs of cardiac tamponade, fever, weight loss and night sweats in a subject having Pericardial tuberculosis.
  • the methods treat or prevent one or more symptoms of Bone and Joint tuberculosis in a subject in need thereof.
  • Tuberculous osteomyelitis can affect all bones including the ribs, skull, phalanx, pelvis, and long bones. Tuberculosis is the most common infectious cause for single of multiple osteomyelitic rib lesions since other causes of osteomyelitis of the rib are rare.
  • the methods treat or prevent one or more of tuberculous arthritis orthopnea, tuberculous osteomyelitis, back pain or stiffness, paralysis, deformity, tenosynovitis of the hand, arthritis of the wrist, and carpal tunnel syndrome in a subject having skeletal tuberculosis.
  • the methods treat or prevent one or more symptoms of miliary tuberculosis in a subject in need thereof.
  • miliary tuberculosis initially was used to describe the resemblance of the pathologic lesions to millet seeds. Now this term denotes any progressive disseminated tuberculosis spread hematogenously. It can be divided into acute miliary tuberculosis, cryptic miliary tuberculosis, and nonreactive tuberculosis.
  • Acute miliary tuberculosis is associated with a brisk and histologically typical tissue reaction. Children have acute or subacute onset, high intermittent fevers, night
  • Pleural effusion, peritonitis, or meningitis occurs in as many as two thirds of cases.
  • the clinical course of young adults is usually more chronic and initially less severe.
  • now older individuals are affected by miliary tuberculosis more frequently, and their underlying illnesses may obscure the diagnosis.
  • Patients usually have nonspecific constitutional symptoms, such as fever, anorexia, weakness, and weight loss. They may have headache due to meningitis, abdominal pain resulting from peritonitis, or pleural pain caused by pleuritis.
  • Patients may have normal white cell count, anemia, hyponatremia, elevation of alkaline phosphatase and transaminases.
  • Fulminant miliary tuberculosis may be associated with severe refractory hypoxemia (adult respiratory distress syndrome) and disseminated intravascular coagulation.
  • Cryptic miliary tuberculosis occurs in older patients with miliary tuberculosis; chest X-rays are normal and tuberculin test results are negative. Patients have a chronic clinical course characterized by mild intermittent fever, anemia, and, ultimately, meningeal involvement preceding death.
  • Nonreactive tuberculosis is very rare and characterized by massive hematogenous dissemination of tubercle bacilli, "non-granulomatous" ("nonreactive") tissue lesions, and often a septic presentation. Patients present with overwhelming sepsis, splenomegaly and subtle diffuse mottling on the chest X-ray. Hematologic abnormalities include leukopenia, thrombocytopenia, anemia, pancytopenia, leukemoid reactions, myelofibrosis, or polycythemia. Disseminated tuberculosis should be considered when pancytopenia is associated with fever and weight loss.
  • the methods treat or prevent one or more symptoms of acute miliary tuberculosis, cryptic miliary tuberculosis, or nonreactive tuberculosis in a subject having miliary tuberculosis.
  • the methods treat or prevent one or more symptoms of Genitourinary tuberculosis in a subject in need thereof.
  • Genitourinary tuberculosis is mostly a disease of middle-aged adults, and the onset is usually insidious. Asymptomatic renal cortical foci may occur in all forms of tuberculosis. In normal hosts, the interval between infection and active renal disease is
  • Genitourinary symptoms of dysuria and gross hematuria are most common. Constitutional symptoms and Pyuria, albuminuria, and hematuria were the most common laboratory abnormalities. Renal tuberculosis may spread to the prostate, seminal vesicles, epididymis, and testis in that order. The usual clinical manifestations are a tender scrotal mass associated with a draining sinus or oligospermia unresponsive to treatment. Genital foci can also result from lymphatic or hematogenous spread and present as a painful testicular or scrotal mass.
  • the methods treat or prevent one or more symptoms of abdominal tuberculosis in a subject in need thereof.
  • Abdominal tuberculosis can affect the gastrointestinal tract, the peritoneum, the liver, and the pancreas. Tuberculosis can involve any gastrointestinal site from the oropharynx to the anus. Patients can present with nonhealing ulcers of the tongue or oropharynx, or nonhealing sockets after tooth extraction. An adjacent caseous node might result in esophageal stricture with obstruction, tracheoesophageal fistula formation, and rare fatal hematemesis from an aortoesophageal fistula. Patients might have ulcerative or hyperplastic lesions in the stomach or gastric outlet obstruction. Duodenal involvement may lead to symptoms of peptic ulcer or obstruction.
  • enteric tuberculosis The most typical site of enteric tuberculosis is the ileocecal area producing symptoms of pain, anorexia, diarrhea, obstruction, hemorrhage, and a palpable mass. Patients with anal tuberculosis might have ulcers, perianal warty growths, and fistulas. Tuberculous peritonitis results from either spread of adjacent tuberculous disease or military tuberculosis.
  • the clinical picture has two types: plastic and serous.
  • the plastic type is less common; characterized by tender abdominal masses and a "doughy abdomen".
  • the serous type presents with ascites often without signs of
  • Tuberculosis is a frequent cause of granulomatous hepatitis with elevated alkaline phosphatase and gamma-glutamyl transpeptidase levels that are out of proportion to bilirubin levels with normal or mildly elevated transaminase levels. Very rarely, tuberculosis granulomatous hepatitis causes jaundice without evidence of extrahepatic tuberculosis (primary tuberculosis of the liver). Focal hepatic tuberculosis describes single or multiple tuberculous abscesses occurring in patients with little natural immunity to tuberculosis and in children.
  • Pancreatic tuberculosis may present with an abscess or a mass involving local nodes and resembling carcinoma. Abdominal lymph nodes may obstruct the biliary tract causing tuberculous ascending cholangitis. b. Infectious Diseases leading to Granulomas
  • compositions and methods treat or prevent formation of granulomas in a subject in need thereof.
  • Granulomas form in response to chronic inflammation. Accordingly, the most common cause of granulomas are infections. Caseating granulomas are formed by infections, such as tuberculosis and fungal infections. Noncaseating granulomas may be formed by an inflammatory condition (e.g., sarcoidosis and Crohn disease), vasculitis, and exposure to foreign objects.
  • an inflammatory condition e.g., sarcoidosis and Crohn disease
  • granulomas are characteristic of certain diseases. Most commonly, in chronic granulomatous disease (CGD), an inherited genetic mutation reduces the ability of white blood cells to kill certain bacteria and fungi, like Staphylococcus aureus and Aspergillus. Individuals with CGD are highly susceptible to infections that lead to granuloma development throughout the body. Similarly, granuloma annulare is a chronic skin disorder characterized by granulomas appearing as small red or yellow bumps in a ring shape on the skin.
  • CGD chronic granulomatous disease
  • granulomatosis with polyangiitis is a rare autoimmune-induced vasculitis characterized by granuloma formation, causing inflammation of the blood vessels (primarily small-sized arteries) and, ultimately, affecting blood flow.
  • Both components trigger the innate immune response with cytokine (TNFa, IL- la, IL-6, IL- 10, IFNy and chemokine CCL2 production.
  • cytokine TNFa, IL- la, IL-6, IL- 10, IFNy and chemokine CCL2 production.
  • Mycobacterium leprae the causative organism of leprosy lives intracellularly in the skin, nasal mucosa and Schwann cells of the peripheral nerves. As a consequence, there is gross thickening of the facial skin, hypopigmentation and loss of sensation to heat, cold and pain.
  • the Gram -ve bacillus Brucella rapidly multiplies in the lymph node and causes probably by the Lipid A component of its endotoxin destruction of the lymphoreticular organs. When it colonizes the heart tissue it can cause fatal endocarditis.
  • the Gram+ve Listeria secretes the exotoxin Listeriolysin O, a P hemolysin that destroys red cells, neutrophils and monocytes.
  • the fungus Histoplasma has tropism for the mucous membrane in the mouth where it causes lesions. All the quoted pathogens can sustain their intra-macrophage survival by subverting the killing machinery of the cells (arrest of the phagolysosomal fusion, disruption of signaling pathways) thereby assuring the chronicity of the infection. It should be pointed out that because of the intimate relationship between host and invading pathogen it is difficult to clearly isolate the pathogen or host-derived factors in the pathogenesis of the diseases.
  • compositions of the described infected tissue targeting peptides to deliver one or more active agents, such as therapeutic agents, to the site of a granuloma in an amount effective to treat or prevent one or more symptoms of the granuloma in a subject are provided.
  • Tuberculosis is a chronic disease of animals caused by a Mycobacterium sp.
  • Mycobacterium bovis can infect practically all mammals, causing a general state of illness, coughing and eventual death. Therefore, in some forms, the subject is a fish, a mammal or an avian. In some forms, the subject is a shellfish.
  • TB Tuberculosis
  • the subject is a poultry bird, such as a chicken, a turkey, a duck or other fowl raised for human consumption.
  • the subject is a farm animal, such as a cow, pig, sheep, goat or rabbit.
  • the subject is a primate, such as a human. Therefore, in some forms, the subject is a human patient.
  • the subject has been medically diagnosed as having an infection with a microorganism, or a disease or disorder associated with an infection with a microorganism by exhibiting clinical (e.g., physical) symptoms of a disease.
  • the methods administer the compositions including infected tissuetargeting molecules to a subject having one or more symptoms of an infection, disease or disorder associated with Mtb.
  • tuberculosis In immunocompetent individuals, approximately 3-10% of those with tuberculous infection will develop tuberculosis in the first 1-2 years after infection and another 5% will develop tuberculosis during their lifetime.
  • the risk of Mtb infection is modified by the age of acquiring infection (e.g. being lowest in the age range of 5 to 9 years) and many other host factors. Exogenous reinfection is thought to be uncommon in immunocompetent persons residing in areas with a low prevalence of tuberculosis, but life-style- related factors and chronic diseases, such as active or passive smoking, nutritional status and diabetes mellitus may significantly affect the risk of reactivation of endogenous infection.
  • the risk of progressing rapidly to infection once infected with M. tuberculosis, the risk of reactivation, and the risk of exogenous reinfection are all increased compared to HIV-1 seronegative persons.
  • the subject is identified as having an increased risk of an infection with Mtb, and/or developing a disease associated with Mtb.
  • the subject has been medically diagnosed as having a latent Mtb infection, but has no symptoms.
  • the subject has a predisposition to Mtb infection, or to be at risk of a disease or disorder associated with Mtb infection by exhibiting clinical (e.g., physical) symptoms, which are indicative of an increased risk or likelihood of developing Mtb infection, or a disease or disorder associated with Mtb infection.
  • the subject is an immunocompromised subject, or subject having one or more factors associated with increased risk of Mtb infection or active tuberculosis, such as active or passive smoking, having poor or minimal nutritional status, and/or having diabetes mellitus.
  • the methods administer pharmaceutical formulations including the infected tissue-targeting molecules or conjugates thereof to a subject prior to
  • the methods administer pharmaceutical formulations including the Mycobacterium tuberculosis binding peptides to a subject via oral administration, or via sub-cutaneous or intravenous injection.
  • the subject is a patient with an underlying disease or disorder that reduces the efficacy of the immune system for preventing Mtb infection and/or onset of disease caused by Mtb in the subject.
  • the subject has previously received therapy for TB.
  • the subject has resurgent TB, for example, following prior treatment.
  • the subject has HIV.
  • the subject has diabetes mellitus.
  • the subject is an active or passive smoker.
  • any of the methods can include one or more steps of diagnosis of a microbial infection, such as a bacterial infection.
  • the methods detect and diagnose a Mycobacterium infection/detection of the presence of Mycobacterium cells in a subject or sample.
  • the compositions of infected tissuetargeting molecules are attached to a dye, or other imaging agent, to label and identify Mycobacterium bacteria and/or tissues infected therewith.
  • methods of using the labelled compositions to detect and diagnose subjects as having Mycobacterium infection include administering a composition to the subject, or to a sample from the subject and detecting the label or imaging agent in an organ of the subject, or in the sample.
  • the methods can include one or more additional tests known in the art for detecting and typing Mycobacterium bacteria.
  • a treatment regimen can include one or multiple administrations of a pharmaceutical formulation including active agents such as antibiotics and having infected tissue-targeting molecules each independently having an amino acid sequence of any of SEQ ID NOs: 1-37, or a functional variant thereof attached thereto as a targeting agent, and formulations thereof for achieving a desired physiological change, including administering to an animal, such as a bird or mammal, especially a human being, an effective amount of the compositions to treat the disease or symptom thereof, or to produce the physiological change.
  • the desired physiological change is the reduction in the amount of pathogenic Mycobacterium bacteria
  • 45652286 in the body i.e., in a bodily location or site, such as the blood and/or tissues and organs of the subject.
  • the treatment regimens include administering the compositions to treat a disease or symptom associated with infection by pathogenic Mycobacterium bacteria in the subject.
  • the methods are administered in a regimen together with other treatments or procedures, for example, to treat and/or prevent pneumonia in the subject.
  • compositions including active agents such as antibiotics and having infected tissue-targeting molecules each independently having an amino acid sequence of any of SEQ ID NOs: 1-37, or a functional variant thereof attached thereto as a targeting agent are typically sufficient to reduce or alleviate one or more symptoms associated with pathogenic Mycobacterium infection in a subject are provided.
  • Symptoms of diseases and disorders associated with pathogenic Mycobacterium infection may be physical, such as fever, or biological such as increased amounts of one or more toxin(s) in the blood.
  • the compositions include nanoparticulate compositions, such as porous silicon nanoparticles loaded with antibiotics and having infected tissue-targeting molecules each independently having an amino acid sequence of any of SEQ ID NOs: 1- 37, or a functional variant thereof attached thereto as a targeting agent.
  • the amount of nanoparticles can be effective to, for example, treat or prevent one or more symptoms of a pathogenic Mycobacterium infection in a subject.
  • the nanoparticles are delivered systemically, for example, orally or via sub-cutaneous or intravenous injection, or by inhalation.
  • the nanoparticulate compositions are non-toxic in vivo and do not target or otherwise modulate non-infected tissue or metabolic products.
  • the nanoparticles are administered in an amount effective to reduce one or more symptoms associated with the pathogenic Mycobacterium infection in the subject. In some forms, the nanoparticle composition is administered in an effective amount to prevent Mycobacterium infection in the subject, or to prevent one or more diseases or disorders associated with pathogenic Mycobacterium infection in a subject at risk of severe disease resulting from pathogenic Mycobacterium infection.
  • the therapeutic result of the nanoparticles can be compared to a control.
  • Suitable controls are known in the art.
  • a typical control is a comparison of a condition or symptom of a subject prior to and after administration of the nanoparticle composition.
  • the condition or symptom can be a biochemical, molecular, physiological, or pathological
  • the effect of the nanoparticle compositions on a particular symptom, pharmacologic, or physiologic indicator can be compared to an untreated subject, or the condition of the subject prior to treatment.
  • the symptom, pharmacologic, or physiologic indicator is measured in an infected subject prior to treatment, and again one or more times after treatment is initiated.
  • the control is a reference level, or average determined based on measuring the symptom, pharmacologic, or physiologic indicator in one or more subjects that do not have the disease or infection to be treated (e.g., healthy subjects).
  • the effect of the treatment is compared to a conventional treatment that is known the art.
  • the actual effective amounts of the nanoparticle composition can vary according to factors including the specific nanoparticles administered, the particular composition formulated, the mode of administration, and the age, weight, condition of the subject being treated, as well as the route of administration and the disease or disorder.
  • the active agent that is administered is an antibiotic. Since some antibiotics, such as vancomycin, are to some extent toxic to a subject when administered systemically at therapeutic amounts, the described compositions including infected tissue-targeting molecules reduce the toxicity of the therapeutically effective dose of the antibiotics by selectively targeting the antibiotics to the site of infection. Therefore, in some forms, the methods reduce the toxicity of the antibiotics required to treat and/or prevent infection with Mtb.
  • the amount of the antibiotics required to treat the infection in the subject is less than the amount of the same antibiotic that is required when administered to a subject alone to treat a corresponding infection. Therefore, in some forms, the amount of the compositions that is required can be compared to a control treated with the same antibiotic and/or same nanoparticles in the absence of the targeting peptide(s).
  • the amount of the antibiotic that is required when loaded into a targeted nanoparticle having Mycobacterium tuberculosis binding peptides each independently having an amino acid sequence of any of SEQ ID NOs:l-37, or a functional variant thereof attached thereto as a targeting agent is about 1/100, 1/50, 1/10, 1/9, 1/8, 1/7, 1/6, 1/5, 14, 1/3, 14 or less than 14 the amount of the same antibiotic that is required to treat the infection when administered in the absence of the nanoparticles.
  • the composition increases the efficacy of the antibiotic by more than 1%, such as by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 75%, 100%, 200%, 300%, 400% or 500%, or more than 500%. In some forms, the composition reduces the
  • 45652286 dosage of the antibiotic that is required to treat or prevent the infection by up to 5%, 10%, 20%, 30%, 40%, 50%, 60%, 75%, 100%, 200%, 300%, 400% or 500%, or more than 500%.
  • the effective amount of nanoparticle compositions causes little or no killing of the cells of the subject, and preferably little or no inhibition of metabolism in the cells. It is particularly preferred that the composition have little or no effect on the normal bacterial flora of the subject outside of the site of the bacterial infection.
  • dosages of nanoparticles are administered once, twice, or three times daily, or every other day, two days, three days, four days, five days, or six days to a human.
  • dosages of nanoparticle compositions are administered about once or twice every week, every two weeks, every three weeks, or every four weeks.
  • dosages are administered about once or twice every month, every two months, every three months, every four months, every five months, or every six months.
  • the regimen includes one or more cycles of a round of therapy with the nanoparticle compositions followed by a drug holiday (e.g., no compositions conjugated or complexed with the infected tissue-targeting molecules nanoparticles).
  • the round of the therapy can be, for example, any of the administrations discussed above.
  • the drug holiday can be 1, 2, 3, 4, 5, 6, or 7 days; or 1, 2, 3, 4 weeks, or 1, 2, 3, 4, 5, or 6 months.
  • the subject is administered a dosage of between about 0.1 mg/kg body weight and 100 mg/kg body weight, inclusive, of the composition, such as nanoparticles having infected tissue binding molecules each independently having an amino acid sequence of any of SEQ ID NOs: 1-37, or a functional variant thereof attached thereto as a targeting agent and having loaded therein an antibiotic such as vancomycin.
  • the subject is administered a dosage of between about 0.1 mg/kg body weight and 1,000 mg/kg body weight, inclusive, of the composition.
  • the subject is administered a dosage of between about 0.2 mg/kg body weight and 10 mg/kg body weight, inclusive, of the composition.
  • the subject is administered a dosage of between about 0.3 mg/kg body weight and 1,000 mg/kg body weight, inclusive, of drug-filled nanoparticles that are conjugated or complexed with the Mycobacterium tuberculosis binding peptides. In some forms, the subject is administered a dosage of between about 0.4 mg/kg body weight and 500 mg/kg body weight, inclusive, of the composition. In some forms, the subject is administered a dosage of between about 0.5 mg/kg body weight and 100 mg/kg body weight, inclusive, of the composition. In some
  • the subject is administered a dosage of between about 1.0 mg/kg body weight and 100 mg/kg body weight, inclusive, of drug-filled nanoparticles that are conjugated or complexed with the infected tissue-targeting molecules.
  • the subject is administered a dosage of between about 1.0 mg/kg body weight and 5 mg/kg body weight, inclusive, of the composition.
  • Particular dosage regimens include, for example, one or more cycles in which the subject is administered the of the composition on each of two, three, four, five, six or seven days, weeks or months in a row, followed by a one, two, three, four, five, six or seven-day, week, or month drug holiday.
  • compositions lead to direct or indirect reduction in the number of viable cells of pathogenic Mycobacterium bacteria in the body (e.g., in the blood or tissue or organs) and/or direct or indirect inhibition of the production of Mycobacterium toxins or equivalents, or the direct or indirect reduction of one or more symptoms of a disease or disorder associated with pathogenic Mtb bacteria in the subject.
  • the effective amount of compositions is suitable for achieving a specific target amount of antibiotic in the body, such as a specific concentration at the site of infection.
  • the composition delivers an effective amount of an antibiotic for achieving therapeutic efficacy against a strain of pathogenic Mycobacterium that is resistant to the antibiotic when administered at a pharmaceutically acceptable dosage in the absence of the composition.
  • the methods administer antimicrobial agents conjugated or complexed with one or more infected tissue-targeting molecules each independently having an amino acid sequence of any one or more of SEQ ID NOs: 1-37, or functional variants thereof to reduce side effects associated with the anti-microbial agents in the subject, for example, by reducing the systemic exposure to the drug and/or dosage of the therapeutically effective amount.
  • the methods administer vancomycin conjugated or complexed with one or more infected tissue-targeting molecules each independently having an amino acid sequence of any one or more of SEQ ID NOs: 1-37, or functional variants thereof to reduce side effects associated with vancomycin, such as local pain, thrombophlebitis, damage to the kidneys (nephrotoxicity) and/or to the hearing (ototoxicity).
  • compositions are administered alone, or in combination with one or more conventional therapies, for example, a conventional antibiotic or other therapy.
  • tuberculosis is treated with at least two drugs to which the isolate is susceptible.
  • the duration of the treatment regimen, expected drug toxicities, and overall effectiveness will depend on the drugs used.
  • drugs approved by the United States Food and Drug Administration (FDA) for treating tuberculosis.
  • FDA United States Food and Drug Administration
  • the antibiotics are two or more of those associated clinically with treatment of Mtb infection, such as first- line oral drugs (with typically adult daily dose) Isoniazid (300 mg), Rifampin (600 mg), Rifabutin (300 mg), Pyrazinamide (30 mg/kg), Ethambutol (15 - 25 mg/kg); and injectable drugs (with typically adult daily dose): Streptomycin (15 mg/kg), Amikacin (15 mg/kg), Kanamycin (15 mg/kg), Capreomycin (15 mg/kg).
  • first- line oral drugs with typically adult daily dose
  • Isoniazid 300 mg
  • Rifampin 600 mg
  • Rifabutin 300 mg
  • Pyrazinamide 30 mg/kg
  • Ethambutol (15 - 25 mg/kg
  • injectable drugs with typically adult daily dose: Streptomycin (15 mg/kg), Amikacin (15 mg/kg), Kanamycin (15 mg/kg), Capreomycin (15 mg/kg).
  • the antibiotics two or more of those associated clinically with treatment of Mtb infection such as Second-line oral drugs (with typically adult daily dose): Bedaquiline (400 qd or 200 mg q.i.d.), Pretomanid (200 mg qd), Linezolid (600 or 1,200 mg qd), Ofloxacin (400 mg b.i.d.), Levofloxacin (500 mg qd), Moxifloxacin (400 mg qd), Gatifloxacin (400 mg qd), Ethionamide (250 mg b.i.d. or t.i.d.) , Aminosalicylic acid (3 g q.i.d.), and Cycloserine (250 mg b.i.d. or t.i.d.).
  • Second-line oral drugs with typically adult daily dose: Bedaquiline (400 qd or 200 mg q.i.d.), Pretomanid (200 mg qd), Linezolid (600 or 1,200 mg
  • the antibiotics are two or more of Isoniazid (300 mg), Rifampin (600 mg), Rifabutin (300 mg), Pyrazinamide (30 mg/kg), Ethambutol (15 - 25 mg/kg); and injectable drugs (with typically adult daily dose) Streptomycin (15 mg/kg), Amikacin (15 mg/kg), Kanamycin (15 mg/kg), Capreomycin (15 mg/kg), Ofloxacin (400 mg b.i.d.), Levofloxacin (500 mg qd), Moxifloxacin (400 mg qd), Gatifloxacin (400 mg qd), Ethionamide (250 mg b.i.d. or t.i.d.), Aminosalicylic acid (3 g q.i.d.), and Cycloserine (250 mg b.i.d. or t.i.d.).
  • the methods include administration of nanoparticles having infected tissue-targeting molecules each independently having an amino acid sequence of any of SEQ ID NOs:l-37, or a functional variant thereof attached thereto as a targeting agent and having loaded therein an antibiotic such as vancomycin in combination with one or more additional active agents.
  • the combination therapies can include administration of the described compositions and other active agents together in the same admixture, or in
  • the pharmaceutical composition includes drug-filled nanoparticles that are conjugated or complexed with the infected tissuetargeting molecules and one, two, three, or more additional active agents.
  • the additional active agent(s) can have the same, or different mechanisms of action.
  • the combination results in an additive effect on the treatment of pathogenic Mycobacterium infection. In some forms, the combinations result in a more than additive effect on the treatment of the disease or disorder.
  • the additional therapy or procedure can be simultaneous or sequential with the combination therapy.
  • the additional therapy is performed between drug cycles or during a drug holiday that is part of the compositions dosage regime.
  • the additional therapy or procedure is surgery.
  • Additional therapeutic agents include conventional therapeutics such as nonsteroidal anti-inflammatory drugs (NSAID), pain relief and antimicrobial drugs. Therefore, in some forms, the additional therapeutic agents include antibacterial antibiotics, bacteriophage and immunotherapies. In some forms, the additional active agent is another drug directly or indirectly targeting pathogenic Mtb, or other microorganisms present or suspected to be present at the site of infection.
  • NSAID nonsteroidal anti-inflammatory drugs
  • the additional active agent is another drug directly or indirectly targeting pathogenic Mtb, or other microorganisms present or suspected to be present at the site of infection.
  • the methods administer two or more antimicrobial agents that are encapsulated alone or together within particles conjugated or complexed with one or more infected tissue-targeting molecules each independently having an amino acid sequence of any one or more of SEQ ID NOs: 1-52, or functional variants thereof.
  • the methods administer vancomycin together with Ceftazidime encapsulated alone or together within particles conjugated or complexed with one or more Mycobacterium tuberculosis binding peptides each independently having an amino acid sequence of any one or more of SEQ ID NOs: 1-52, or functional variants thereof.
  • a method for treatment or prevention of an infection by a microbial pathogen in a subject in need thereof including administering to the subject a pharmaceutical formulation including:
  • the antimicrobial agent is in an amount effective to treat or prevent one or more symptoms of the infection in the subject.
  • a method for identification of an infection by a microbial pathogen in a subject including administering to the subject a pharmaceutical formulation including:
  • an imaging or diagnostic agent wherein the imaging or diagnostic agent is in an amount effective to bind to infected tissues and/or microbial pathogens, and label or otherwise identify the infection in the subject.
  • infected tissue homing peptide includes any one or more of SEQ ID NOs:l-52, or a functional variant thereof having at least 70% sequence identity to any one of SEQ ID NOs:l-37.
  • infected tissue homing peptide includes a functional variant having at least 70% sequence identity to PPRRGLIKLKTS (SEQ ID NO:1).
  • the infected tissue homing peptide includes a functional variant having at least 70% sequence identity to GRPARPAR (SEQ ID NO:3).
  • infected tissue homing peptide includes a functional variant having at least 70% sequence identity to GRP ARP AR (SEQ ID NO:3).
  • the infected tissue homing peptide includes a functional variant having at least 70% sequence identity to AKRGARSTA (SEQ ID NO:5).
  • the formulation further includes one or more polypeptides or other amino acid sequences, wherein the polypeptides or other amino acid sequences are contiguous with the amino (NH) or carboxyl (COOH) terminus of the infected tissue homing peptide.
  • the formulation further includes one or more additional molecules selected from the group including a carbohydrate, a lipid, a small organic molecule, a polymer, and a nucleic acid.
  • the antimicrobial agent is selected from the group including bedaquiline, pretomanid, Rinzolid, delamanid, isoniazid, vancomycin, rifampin, rifabutin, pyrazinamide, ethambutol, streptomycin, amikacin, kanamycin, capreomycin, ofloxacin, levofloxacin, moxifloxacin, gatifloxacin, ethionamide, aminosalicylic acid, and cycloserine.
  • the delivery vehicle is selected from the group including a citric acid silver nanoparticle, a liposome, a micelle, a viral capsid, a polymeric particle, a dendrimer, a porous silicon particle, a metal particle, and an iron oxide nanoparticle.
  • the Mycobacterium sp. is resistant to one or more antibiotics selected from the group including bedaquiline, pretomanid, Rinzolid, delamanid, isoniazid, vancomycin, rifampin, rifabutin, pyrazinamide, ethambutol, streptomycin, amikacin, kanamycin, capreomycin, ofloxacin, levofloxacin, moxifloxacin, gatifloxacin, ethionamide, aminosalicylic acid, and cycloserine.
  • antibiotics selected from the group including bedaquiline, pretomanid, Rinzolid, delamanid, isoniazid, vancomycin, rifampin, rifabutin, pyrazinamide, ethambutol, streptomycin, amikacin, kanamycin, capreomycin, ofloxacin, levofloxacin, moxifloxacin,
  • M. tuberculosis (Mtb) infection selected from the group including fever, chest pain, wheezing or difficulty breathing, cough, fatigue, and unintentional weight loss.
  • 45652286 (b) an antimicrobial agent, wherein the antimicrobial agent is in an amount effective to treat or prevent one or more symptoms of the infection in the subject.
  • Example 1 M. tuberculosis-binding peptides that penetrate the cellular rim and/or the caseous core of TB lesions identified by peptide library screening
  • peptide receptors in M. tuberculosis-infected lung vs control lung tissue were stained.
  • the readout is not quantitative, but highly specific staining of Mtb-infected tissues with antibodies to known peptide receptors can identify key receptors for further studies.
  • Antibodies against receptors p32, NRP-1, Fn-EDB and TNC-C were made inhouse, the rest of the required antibodies were ordered from commercial companies. Fluorescent immunostainings of both necrotic and cellular lesions were imaged (at least 3 animals and 7 images per tissue) with Olympus Fluoview FV1200 MPE confocal microscope.
  • Table 1 List of the antibodies against peptide receptors used in the immunostaining analysis.
  • Example 2 In vivo playoff auditioning of the candidate peptides for targeting the TB lesions
  • In vivo playoff auditioning is useful for head-to-head comparative evaluation of multiple peptide phages in the same animal.
  • the most important advantage of in vivo playoff studies - that all the peptide phages are auditioned in parallel under identical conditions - avoids inter-animal variability that may arise due to differences in physiological state of the animal and/or disease status heterogeneity and subtle variations in experimental conditions.
  • inclusion of positive and negative phages provides quality control for homing experiments.
  • Some peptides that have been shown in the literature to specifically home and penetrate tumors may also recognize infected tissue and could potentially be used in targeted delivery of treatments to bacterial infections.
  • the aim was to test such already available peptides to identify ones that accumulate in TB-infected tissues, and that therefore can potentially be used to deliver therapeutic agents to the sites of infection.
  • a significant advantage of testing these peptides first is that they are already at hand.
  • selectively concentrating an antibiotic in macrophages may increase the efficacy of bacterial eradication because macrophages are an important reservoir of intracellular bacilli.
  • phage library screening in vivo play off platform to identify peptides that recognize the bacilli rather than tissue changes brought about by the infection.
  • Such peptides can take a drug payload all the way to the bacilli, not just into the infected area in general.
  • These screens will include an in vivo screening step, and may also yield peptides that recognize new tissue changes caused by the infection, similar in principle to the targets recognized by existing peptides, but distinct from them.
  • Granuloma tissue in particular is likely to upregulate molecules that are not expressed in normal tissues at significant levels and may present new targets detectable by phage display.
  • a pool of equally represented phages displaying candidate and control peptides was administered to live mice (Mtb-infected and healthy uninfected), and the representation of each phage in target and control organs was determined by sequencing (Sanger sequencing for low complexity pools and HTS for high complexity pools).
  • a phage play-off pool with 23 peptide-phages was prepared (encompassing the candidate homing peptides listed in Table 2). This required cloning of some of the peptides, sequencing of all the peptides with Sanger method, amplification, purification and determining the titer of the samples. TB-infected and non-infected mice were dosed with the phage pool and DNA samples of lungs and livers were sent for high throughput sequencing and data analysis.
  • PL 1 peptide (PPRRGLIKLKTS ; SEQ ID NO : 1 ) showed the highest difference between infected and non-infected lung tissue (Figure 2B) and highest fold over the control peptide in infected lungs ( Figure 2A); PL1 interacts with receptors Fn- EDB and TNC-C that are both present in the solid tumor extracellular matrix, and absent in normal adult tissues (Lingasamy et al., 2019). PL 1 has been shown to target also endometriotic lesions (Simon-Gracia et al., 2021).
  • Table 2 Components of in vivo play-off screen and peptides used in the phage pool dosed in TB mice. over the control peptide in both infected and uninfected lungs in the play-off screen
  • RPARPAR peptide (GRPARPAR; SEQ ID NOG) showed high presence in infected and normal lung tissue ( Figure 2A).
  • RPARPAR a prototypic C-end rule (CendR) peptide interacts with NRP1 protein and accumulates in the lung parenchyma when injected intravenously to mice (Teesalu et al., 2009).
  • IP3 peptide contains a hyaluronan-binding motif that homes to macrophage-rich regions in peritoneal tumors (gastric and colon tumors), including poorly vascularized micro-tumors (Ikemoto et al., 2017).
  • LinTTl peptide (AKRGARSTA; SEQ ID NOG) had 3 times higher fold in infected lungs over the non-infected lungs ( Figure 2B). LinTTl is targeting activated macrophages in the tumors and inflammatory lesions (Sharma et al., 2017).
  • Nanoparticles identified from the screens were tested for their ability to deliver drugs to the sites of infection at concentrations and depth of penetration not achieved in conventional therapy. The delivery may be best accomplished with a drug-filled nanoparticle coated with a TB homing peptide for infection targeting. Nanoparticles can be designed to release the drug slowly for a long-lasting therapeutics effect, which would reduce the frequency of treatments. Nanoparticles (or small dendrimers) have significant advantages over simple peptide-drug conjugates as drug delivery vehicles, such as increasing the efficacy of the peptide. The presentation of the peptide at the surface of a nanoparticle allows for multivalent binding that makes up for the fact that peptides generally do not have high affinities for their target (Ruoslahti, 2012).
  • Nanoparticles can be designed to have a favorable drug load to target molecule ratio.
  • peptide-coated nanoparticles have already been used in bacterial targeting and can readily be redesigned for TB targeting.
  • 45652286 reproduce the tissue-penetrating and cell-penetrating properties of the peptides, and the drug payload to target molecule ratio is less favorable than with nanoparticles.
  • Citric AgNP have an average core size of 62 ⁇ 20 nm and hydrodynamic size of 103 ⁇ 40 nm ( Figures 3A-3C) (Lingasamy et al., 2020).
  • AgNPs were developed as a model platform for targeted in vitro and in vivo delivery (Tobi et al., 2021).
  • the AgNPs have several unique features that make them useful for biodistribution studies including plasmonic enhancement emission from coupled fluorescent dyes to allow ultrasensitive imaging of single nanoparticles.
  • Example 4 Evaluation of priority peptides that were identified in an initial in vivo biopanning
  • the drawback of the standard mouse infection models for tuberculosis is their lack of advanced lung lesion types as the progression of disease rarely reaches stages of extensive necrosis and calcification in lungs.
  • the C3HeB/FeJ mouse strain with a recessive allele was studied first by Igor Kramnik, et al (2000).
  • the C3HeB/FeJ TB efficacy model has been optimized for several years now at CSU in terms of achieving consistent infections and reproducibly showing caseous necrotic lesion types at treatment start.
  • the model uses a low dose aerosol infection with M. tuberculosis Erdman (50-120 CFU/ mouse), with treatment start at 8 weeks post aerosol.
  • the lung pathology in C3HeB/FeJ mice is heterogeneous, generating diverse lesion types which present unique microenvironments each harboring metabolically distinct bacterial subpopulations (Driver et al., 2012; Irwin et al., 2015).
  • the C3HeB/FeJ mouse infection model is characterized by developing hypoxic, caseous necrotic lesions in lungs upon an M. tuberculosis infection.
  • lesion types Three distinct lesion types have been characterized, including highly encapsulated caseous necrotic granulomas (type I), fulminant neutrophilic alveolitis (type II), and cellular non-necrotizing lesions (type III), with only the latter lesion type being present in BALB/c mice.
  • These different lesion types exhibit highly different microenvironmental conditions that vary in certain parameters such as oxygen tension, pH, nutrient supply, and cellular composition and are more closely aligned with the heterogamous pathology found in human TB patients (Lenaerts et al., 2015).
  • caseum with a higher ratio of extracellular to intracellular bacilli, and the altered microenvironments that arise, are known to have a profound impact on drug efficacy.
  • Synthetic fluorescein-labelled peptides and peptide-functionalized silver nanoparticles loaded with CF555 (AgNP) were administered to Mtb Erdman infected C3HeB/FeJ mice as a single intravenous (IV) or intraperitoneal (IP) injection and allowed to circulate as indicated prior to a full body perfusion to remove unbound material (Figure 4).
  • Table 3 Synthetic fluorescein-labelled peptides and peptide-functionalized silver nanoparticles loaded with CF555 dose groups.
  • a panel of existing homing peptides was tested for ability to home to TB infected tissues using a series of experiments of increasing labor intensity.
  • In vivo playoff was carried out using a set of ⁇ 20 phages displaying homing peptides with targeting specificity towards activated macrophages, angiogenic sites, and activated extracellular matrix.
  • TB infected mice were injected with equimolar mixture of different candidate and control peptide phages, and their representation in TB lesions and control tissues will be studied by high-throughput sequencing.
  • the expression of receptors for the peptides showing systemic selectivity towards TB lesions was studied by immunostaining to confirm the playoff data and to develop better understanding of the peptide target sites.
  • systemic homing experiments with selected fluorophore-labeled synthetic peptides were performed in TB infected mice, and peptide bio-distribution tested by confocal immunofluorescence imaging.
  • tissue sections from TB-infected tissue with antibodies against the known target molecules (receptors) identified the lead peptides.
  • tissue sections stained with markers of vascular endothelial cells and other cell types in the lesion with existing materials and with no exposure to M. tuberculosis bacteria provided information on the presence and spatial distribution of peptide receptors to guide decisions as to which peptides to test further.

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Abstract

Des polypeptides qui ciblent sélectivement un tissu infecté et/ou des pathogènes microbiens in vivo et qui s'y lient ont été découverts. Des agents antimicrobiens et des agents de diagnostic tels que des colorants qui sont conjugués aux molécules de localisation de tissu infecté sont efficacement ciblés sélectivement sur des tissus infectés et/ou des pathogènes microbiens in vivo. L'invention concerne des compositions et des procédés de molécules de liaison de tissu infecté pour la localisation sélective de tissus infectés. Dans certains modes de réalisation, les compositions comprennent des nanoparticules, telles que des nanoparticules d'argent, conjuguées aux peptides de localisation de tissu infecté et chargées d'agents antimicrobiens. Généralement, la quantité d'antimicrobien qui est nécessaire pour traiter ou prévenir une infection par des pathogènes tels que Mycobacterium tuberculosis (Mtb) lorsqu'elle est administrée dans les nanoparticules est inférieure à la quantité du même antimicrobien administré seul.
PCT/US2024/031548 2023-05-31 2024-05-30 Molécules ciblant des tissus infectés et leurs procédés d'utilisation Pending WO2024249553A1 (fr)

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