Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
  • A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/66—Phosphorus compounds
  • A61K31/662—Phosphorus acids or esters thereof having P—C bonds, e.g. foscarnet, trichlorfon
  • A61K31/663—Compounds having two or more phosphorus acid groups or esters thereof, e.g. clodronic acid, pamidronic acid
• • 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
  • A61K47/54—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 an organic compound
  • A61K47/548—Phosphates or phosphonates, e.g. bone-seeking
• • 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
  • A61K47/54—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 an organic compound
  • A61K47/55—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 an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug
  • A61K47/552—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 an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug one of the codrug's components being an antibiotic
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P19/00—Drugs for skeletal disorders
  • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
• • 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
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07J—STEROIDS
  • C07J51/00—Normal steroids with unmodified cyclopenta(a)hydrophenanthrene skeleton not provided for in groups C07J1/00 - C07J43/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
  • A61B17/00—Surgical instruments, devices or methods
  • A61B2017/00831—Material properties
  • A61B2017/00889—Material properties antimicrobial, disinfectant

Definitions

• the present disclosure relates to antibiotic compounds and, in particular, antibiotic compounds which bind to bone and are useful in the treatment of bone infections.
• Treating bone infections is complicated by their relatively low vascularization and their location beneath soft tissue in the body.
• High doses of systemic drugs and long treatment regimens may be required to provide effective concentrations of the drug at the site of infection.
• this can cause harmful side effects and thereby limit the use of certain antibiotics.
• While local delivery of antibiotics to surgical sites has been explored, this approach is still in the experimental phase and requires an open surgical site.
• widespread antibiotic use over long periods of time has resulted in the development of antibiotic resistant strains of bacteria.
• the present disclosure relates to antibiotic compounds, and in particular, antibiotic compounds which bind to bone and are useful in the treatment of bone infections.
• the inventors conjugated a cationic steroid antimicrobial (CSA) to a bone-binding moiety via a linker.
• CSA cationic steroid antimicrobial
• the bone-binding moiety targets the CSA moiety to the bone, thereby increasing its concentration at the site of a bone infection for both treatment of existing infection and prophylaxis against subsequent infections or in cases of increased risk of infection.
• B is a bone-binding moiety
• L is a linker
• C is a cationic steroid antimicrobial (CSA) moiety.
• the CSA is selected from the group consisting of CSA-8, CSA- 13, CSA-44, CSA-90, CSA-91, CSA-124, CSA-131, CSA-133, CSA-138, CSA- 142, CSA-144, CSA-190, CSA-191, and CSA-192.
• the CSA is CSA-90.
• the bone-binding moiety is a bisphosphonate, such as a bisphosphonate selected from the group consisting of etidronate, clodronate, tiludronate, pamidronate, medronate, etidronate, neridronate, olpadronate, alendronate, ibandronate, aminomethylene diphosphonate, risedronate and zoledronate.
• the bisphosphonate is selected from the group consisting of alendronate, pamidronate and neridronate.
• the bisphosphonate is alendronate.
• the linker is hydrophilic. In some embodiments, the linker has a molecular weight of less than about 2 kDa. In some embodiments, the linker comprises polyethylene glycol (PEG), such as where the linker has a structure of Formula (II):
• X is independently selected from O and S;
• T is absent or is an alkanediyl group having between 1 and 15 carbon atoms
• Y is absent or is an alkanediyl group having from 1 to 15 carbon atoms
• n is an integer from 1 to 30, and the squiggly lines represent points of attachment to the CSA and bone-binding moieties.
• X is O.
• T is an alkanediyl group having from 1 to 15 carbon atoms.
• Y is an alkanediyl group having from 1 to 15 carbon atoms.
• T is an alkanediyl group having from 1 to 6 carbon atoms.
• Y is an alkanediyl group having from 1 to 6 carbon atoms.
• n is an integer from 10 to 20.
• the compound has a structure of Formula (III):
• n is from 1 to 50. In some embodiments, n is from 1 to 30. In some embodiments, the compound has a molecular weight of from about 1.5 kDa to about 2.5 kDa.
• the compound has a structure of Formula (IV):
• compositions comprising a compound disclosed herein and a pharmaceutically acceptable carrier.
• the composition is suitable for systemic administration.
• the composition is suitable for oral administration or parenteral administration.
• the composition is suitable for intravenous administration.
• the infection is a bacterial infection, such as a Staphylococcus aureus infection, a Staphylococcus epidermidis infection, or a Pseudomonas aeruginosa infection.
• the bone comprises a fracture.
• osteomyelitis in a subject, comprising administering to the subject a compound or a pharmaceutical composition disclosed herein.
• the osteomyelitis is associated with a Staphylococcus aureus infection, a Staphylococcus epidermidis infection, or a Pseudomonas aeruginosa infection.
• the subject suffers from a bone disorder selected from the group consisting of a bone fracture, spinal cord injury, spinal disc degeneration, Paget’s disease, bone cancer, metastatic bone cancer, and osteoporosis.
• a bone of the subject is infected with one or more species of bacteria, such as one or more of Staphylococcus aureus, Staphylococcus epidermidis, or Pseudomonas aeruginosa.
• the compound or the pharmaceutical composition is administered systemically to the subject. In some embodiments, the compound or the pharmaceutical composition is administered orally or parenterally to the subject. In some embodiments, the compound or the pharmaceutical composition is administered intravenously to the subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.
• FIG. 1A-C HPLC profiles of BBA-1 ( Figure 1A), pure NHS-PEG-COOH linker ( Figure IB), and pure CSA-90 ( Figure 1C).
• Figure 2A-D FT-IR spectra of alendronate (Figure 2A), CSA- 90 ( Figure 2B), NHS-PEG-COOH linker (Figure 2C), and BBA-1 ( Figure 2D).
• Figure 3A 3 ⁇ 4 NMR spectra of BBA-1, NHS-PEG-COOH linker, CSA-90, and alendronate.
• Figure 3B 31 P NMR spectra of BBA-1 and alendronate.
• Figure 4A-C Figure 4A shows a Kirby Bauer assay against S. aureus and MRSA (results are from a single experiment containing triplicates of each sample).
• Figure 4B shows representative image of zone of inhibition (Kirby Bauer assay against S. aureus and MRSA).
• Figure 4C shows MIC and MBC of CSA-90 and BBA-1.
• Figure 5A-B HA binding assays.
• Figure 8 Data from toxicity study involving BBA- 1 administered to mice showing infection relating to swab assay of soft tissue.
• Figure 9 Data from the toxicity study involving BBA-1 administered to mice showing infection relating to pin assay.
• the articles“a” and“an” are used herein to refer to one or to more than one (/. e. to at least one) of the grammatical object of the article.
• the term“about” is understood to refer to a range of ⁇ 10%, such as ⁇ 5%, or ⁇ 1%, or ⁇ 0.1%.
• administration concurrently or“administering concurrently” or“co administering” and the like refer to the administration of a single composition containing two or more actives, or the administration of each active as separate compositions and/or delivered by separate routes either contemporaneously or simultaneously or sequentially within a short enough period of time that the effective result is equivalent to that obtained when all such actives are administered as a single composition.
• imultaneously is meant that the active agents are administered at substantially the same time, and preferably together in the same formulation.
• pharmaceutically acceptable refers to substances that do not cause substantial adverse allergic or immunological reactions when administered to a subject.
• A“pharmaceutically acceptable carrier” includes, but is not limited to, solvents, coatings, dispersion agents, wetting agents, isotonic and absorption delaying agents, and disintegrants.
• prevention includes reduction of risk, incidence and/or severity of a condition or disorder.
• treatment and“treat” include both prophylactic or preventive treatment (that prevent and/or slow the development of a targeted pathologic condition or disorder) and curative, therapeutic or disease-modifying treatment, including therapeutic measures that cure, slow down, lessen symptoms of, and/or halt progression of a pathologic condition or disorder; and treatment of patients at risk of contracting a disease or suspected to have contracted a disease, as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition.
• the terms“treatment” and“treat” do not necessarily imply that a subject is treated until total recovery.
• treatment also refer to the maintenance and/or promotion of health in an individual not suffering from a disease but who may be susceptible to the development of an unhealthy condition.
• treatment and “treat” are also intended to include the potentiation or otherwise enhancement of one or more primary prophylactic or therapeutic measures.
• a treatment can be performed by a patient, a caregiver, a doctor, a nurse, or another healthcare professional.
• alkanediyl is understood to refer to a bivalent saturated branched or straight chain hydrocarbon group conforming to the formula CnThn.
• Cationic steroid antimicrobials are synthetic compounds designed to mimic the activities of endogenous antibacterial peptides. They are typically cationic and have a broad spectrum of antimicrobial activity including activity against bacteria, fungi and viruses, as well as anti-inflammatory and immunomodulatory activity. More than 100 CSAs have been synthesised.
• Bone is a dynamic tissue and its homeostasis represents a balance between bone formation and bone resorption.
• adult stem cells differentiate into bone progenitor cells (i.e., osteoprogenitor cells) that have the ability to mature into osteoblasts, osteocytes, and form mature bone and mineralized matrix.
• osteoclasts cells that resorb bone tissue dissolve the mineralized matrix and create cavities on the bone surface.
• the compounds described herein may be useful for promoting bone formation and/or treating bone disorders.
• CSA-90 is a small synthetic peptidomimetic compound based on endogenous cationic antibacterial peptides such as the human cathelicidin LL-37.
• LL-37 is found in the airway mucus and is thought to play an important role in controlling bacterial growth in the lung.
• the steroid- like structure of CSA-90 enables it to disrupt cell membranes and therefore confer a broad activity against Gram-positive and Gram-negative bacteria, including vancomycin- and methicillin-resistant strains.
• the compounds described herein comprise a CSA moiety conjugated to a bone binding moiety via a linker.
• the CSA moiety may be any suitable CSA such as CSA-8, CSA-13, CSA-44, CSA-90, CSA-91, CSA-124, CSA-131, CSA-133, CSA-138, CSA-
• the CSA is selected from the group consisting of:
• the CSA moiety is CSA-13, CSA-90, or CSA-131. In certain examples, the CSA moiety is CSA-90.
• the CS A moiety may be attached to the linker via an amino group of a CS A moiety or conjugated to the alkyl side of a CSA molecule.
• a bone-binding moiety as described herein is a chemical group that binds to bone, thereby targeting the compounds of the present disclosure to the bone.
• the bone-binding capability of a particular moiety can be assayed in anumber of different ways.
• bone binding can be assayed by binding to hydroxyapatite (HA), a mineral that is the main inorganic constituent of bone.
• HA hydroxyapatite
• a hydroxyapatite (HA) affinity assay may be performed by incubating a compound conjugated to the moiety in: i) water; and ii) water comprising HA.
• the amount of compound detected in the aqueous phase of the HA solution is expected to decrease as the compound will bind or absorb to the HA surface (See, e.g., Example 4).
• the compound lacking the bone-binding moiety can be assayed in the same way.
• Those skilled in the art will be familiar with other methods by which the bone-binding capability of a particular moiety may be assessed.
• Suitable bone-binding moieties for use in the present disclosure may include, for example, a polyhydoxy-containing moiety, tetracycline derivative, acidic amino acid or peptide, hydoxylated hetercycleo, monophosphonate, bisphosphonate, antibody, or antigen-binding fragment.
• the bone-binding moiety is a bisphosphonate.
• Bisphosphonates generally comprise a phosphate-carbon-phosphate backbone and bind to hydroxyapatite (HA), the major mineral component found in bone, by coordination between the phosphate groups of the bisphosphonate and the calcium ions in the HA.
• the bisphosphonate may be attached to the linker via its terminal functional group (amine group in amino-bisphosphonates), geminal carbon or via either of its phosphate groups.
• the linker is attached to the bisphosphonate via a terminal functional group attached to the geminal carbon of the bisphosphonate, such as for example an amino group or a hydroxy group, or other reactive group.
• bisphosphonate bone binding moieties may include etidronate, clodronate, tiludronate, pamidronate, medronate, etidronate, neridronate, olpadronate, alendronate, ibandronate, aminomethylene diphosphonate, risedronate and zoledronate.
• the bone-binding moiety is alendronate.
• linker described herein covalently attaches a CSA moiety to a bone-binding moiety as set forth in Formula (I):
• B is a bone-binding moiety
• L is a linker
• C is a CSA.
• Formula (I) is not directional and may equally be represented as C-L-B.
• the linker may be small, consisting of a single covalent bond, or it may be larger moiety reaching 10 kDa or more.
• the linker has a molecular weight of less than about 10 kDa, such as less than about 9 kDa, or less than about 8 kDa, or less than about 7 kDa, or less than about 6 kDa, or less than about 5 kDa, or less than about 4 kDa, or less than about 3 kDa, or less than about 2 kDa, such as about 1 kDa or less.
• the linker is from about 0.2 kDa to 10 kDa, such as about 0.3 kDa to 9 kDa, or about 0.4 kDa to 8 kDa, or about 0.5 kDa to 7 kDa, or about 0.5 kDa to 6 kDa, or about 0.5 kDa to 5 kDa, or about 0.5 kDa to 4 kDa, or about 0.5 kDa to 3 kDa, or about 0.5 kDa to 2 kDa, or about 0.5 kDa to 1.5 kDa.
• the linker may include, for example, an ether, ester, thioester, phosphoester, amide, peptide (e.g., dipeptide), polypeptide, polysaccharide, hydrophobic linker (e.g., strait alkane, fatty acid, etc.), or any combination thereof.
• the linker may be a cleavable linker, such as a hydrolysable linker (e.g., carbamate linker) or cathepsin-sensitive linker, or non-cleavable linker.
• the linker is preferably stable in the blood stream for a sufficient period of time (e.g., more than 1 hour, such as more than 6 hours, or more than 12 hours, or more than 18 hours, or more than 24 hours) to enable the compound to reach the bone.
• the linker may be hydrophilic.
• the linker is, or comprises, polyethylene glycol (PEG).
• PEG linker may have a molecular weight as defined in the preceding paragraph.
• the linker has a structure of Formula (II):
• X is independently selected from O and S;
• T is absent or is an alkanediyl group having from 1 to 15 carbon atoms
• Y is absent or is an alkanediyl group having from 1 to 15 carbon atoms
• n is an integer between 1 and 30;
• the squiggly lines represent points of attachment to the CSA and bone-binding moieties.
• T may be an alkanediyl group having from 1 to 12 carbon atoms, or 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to 5 carbon atoms, or 1 to 4 carbon atoms.
• Y may be an alkanediyl group having from 1 to 12 carbon atoms, or 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to 5 carbon atoms, or 1 to 4 carbon atoms.
• T and Y are -CH2CH2-.
• n may be an integer from about 2 to about 25, or from about 2 to about 25, or from about 3 to about 25, or from about 4 to about 25, or from about 5 to about 25, or from about 6 to about 25, or from about 2 to about 20, or from about 2 to about 20, or from about 3 to about 20, or from about 4 to about 20, or from about 5 to about 20, or from about 6 to about 20, or from about 10 to about 30, or from about 10 to about 25, or from about 10 to about 20, or from about 15 to about 25, or from about 12 to about 20.
• the linker may be attached to the CSA moiety via an amino group of the CSA. Likewise, the linker may be atached to the bone-binding moiety via an amino group of the bone-binding moiety.
• the compound represented by Formula (I) may comprise other features, such as further conjugates or moieties.
• the compound may comprise a second antibiotic moiety in addition to the CSA.
• the CSA may be directly conjugated to the second antibiotic moiety, for example, in the form B-L-C-A2, wherein A2 is a second antibiotic moiety.
• the CSA and the second antibiotic moiety may be attached at opposite ends of the compound, for example, in the form A2-B-L-C.
• Suitable second antibiotics may include ciprofloxacin, gemcitabine, paclitaxel, cytarabine, rifalazil, norfloxacin, enoxacin, gatifloxacin, moxifloxacin, a fluoroquinolone ester, a benzoxazinorifamycine, aminoglycoside, polyene, nitroimidazole, rifamycin, bacitracin, a beta-lactam, cephalosporin, chloramphenicol, a glycopeptide, a macrolide, a lincosamide, penicillin, a quinolones, rifampicin, tetracycline, trimethoprim a sulfonamide, amoxicillin, augmentin, amoxicillin, ampicillin, azlocillin, flucloxacillin, mezlocillin, methicillin, cephalexin, cefazedone, cefuroxi
• the compound may comprise more than one bone-binding moiety.
• the bone-binding moieties may be directly attached to each other, such as in the form Bi- B2-L-C, or they may be attached at opposite ends of the compound, such as in the form B1-L-C-B2, wherein Bi and B2 are the same or different bone-binding moieties.
• Bi and B2 are the same or different bone-binding moieties.
• the present disclosure provides a compound having the structure set forth as Formula (III) (BBA-1):
• n is from 1 to 100.
• n is from 1 to 95, or from 1 to 90, or from 1 to 85, or from 1 to 80, or from 1 to 75, or from 1 to 70, or from 1 to 65, or from 1 to 60, or from 1 to 55, or from 1 to 50, or from 1 to 45, or from 1 to 40, or from 1 to 35, or from 1 to 30, or from 5 to 30, or from 5 to 25, or from 5 to 20, or from 10 to 20.
• the compound may have a molecular weight of from about 1 kDato 10 kDa, such as from about 1 kDa to 9 kDa, or from about 1 kDa to 8 kDa, or from about 1 kDa to 7 kDa, or from about 1 kDa to 6 kDa, or from about 1 kDa to 5 kDa, or from about 1 kDa to 4 kDa, or from about 1 kDa to 3 kDa, or from about 1.5 kDa to 3 kDa, or from about 1.5 kDa to 2.5 kDa, or from about 1.75 kDa to 2.25 kDa, such as about 2 kDa.
• the present disclosure provides a compound having the structure set fourth as Formula (IV) (BBA-2):
• the compounds of Formula (I) may, for example, be prepared by reacting an appropriately functionalised linker with a CSA and a bone-binding moiety.
• an appropriately functionalised linker with a CSA and a bone-binding moiety.
• Scheme 1 Exemplary synthesis of a compound of Formula (I)
• a bone-binding moiety having an amino group is reacted with a linker functionalized with two terminal carboxylic acid groups (in which one carboxylic acid group may be activated) so as to provide an intermediate in which the bone-binding moiety is attached to the linker via an amide bond.
• the carboxylic acid present in the intermediate is then reacted with an amino group of a CSA to generate another amide bond and thereby provide the compound of Formula (I).
• the compounds described herein target bone by way of their bone-binding moiety and may be used to treat an infection of a bone.
• the infection may be a bacterial infection such as a Gram-positive bacterial infection or a Gram-negative bacterial infection.
• the bacterial infection is a staphylococcus or pseudomonas infection, such as a Staphylococcus aureus, Staphylococcus epidermidis, or Pseudomonas aeruginosa infection, such as a methicillin-resistant Staphylococcus aureus, methicillin-resistant Staphylococcus epidermidis, or tobramycin-resistant Pseudomonas aeruginosa infection.
• the bacterial infection is a vancomycin-resistant Staphylococcus aureus. Although Staphylococcus epidermidis is sometimes considered a less virulent pathogen than Staphylococcus aureus, reports indicate that Staphylococcus epidermidis strains may be acquiring more invasive properties and are just as effective at forming biofilms, particularly on orthopedic implants (Gill et al. J Bacteriol. 2005. 187: 2426- 2438).
• the bacterial infection is an Aggregatibacter actinomycetemcomitans infection. Aggregatibacter actinomycetemcomitans is commonly observed in cases of jawbone osteomyelitis.
• the bone infection is a fungal infection, such as an infection with a Candida species.
• the infections result from an orthopedic implant, osteomyelitis, or surgical site infection.
• the disclosed compounds are useful for the treatment of infections and, in other embodiments, the disclosed compounds are useful for the prophylaxis of infection in subjects susceptible to, or at risk of, bone infections.
• CSAs like CSA-13, CSA-90, and CSA-131 have also been reported to promote or enhance osteogenesis (US Patent No. 9,694,019; Schindeler et al. J Bone Joint SurgAm. 2015. 97(4): 302-309).
• the compounds described herein may be used to promote bone formation in a subject.
• Bone disorders that may be treated in accordance with the present disclosure include, for example, a bone fracture, a spinal cord injury, spinal disc degeneration, Paget’s disease, bone cancer, and osteoporosis.
• Bone fractures that may be treated using the compositions and methods of the present disclosure include non-union fractures, simple fractures, greenstick fractures, compound fractures, comminuted (multi-fragmentary) fractures, impacted fractures, complicated fractures, hairline fractures, compression fractures, fatigue fractures, and/or pathological fractures.
• Examples of bone fractures that may be advantageously treated by the methods described herein include, but are not limited to, fractures of the spine, leg, and arm.
• a further example of a fracture that may be advantageously treated in accordance with the present disclosure is a vertebral compression fracture.
• Such fracture occurs when one or more of the bones of the vertebral column fractures or collapses, typically when the vertebrae are already weakened for instance as a result of ageing or a disease that weakens bone, such as osteoporosis, Paget’s disease, or bone cancer.
• the bone diseases or conditions that may be treated in accordance with the present disclosure include bone resorption, osteoarthritis, osteoporosis, osteomalacia, osteitis fibrosa cystica, osteochondritis dissecans, osteomalacia, osteomyelitis, osteoblastogenesis, osteopenia, osteonecrosis, and porotic hyperostosis.
• compositions and methods of the present disclosure may be used to treat a subject suffering from an imbalance in bone formation and resorption.
• Imbalance of bone formation and resorption usually causes loss of bone mass and can lead to bone related diseases, such as osteoporosis, rickets, and osteomalacia. These bone diseases are associated with increased risk of bone fractures, increased severity of fractures and protracted time periods for healing. Additionally, with age or injury the incidence of disc degenerative disease or deformity of the spine is increased, leading to spondylolisthesis.
• Dosages may vary with the type and severity of the condition to be treated and may include single or multiple dosses. Specific dosage regimens may be adj usted over time according to the individual need and the professional judgment of the practitioner administering the compound.
• the dosage regimen When administered to a human subject, the dosage regimen may vary depending on a variety of factors including the type and severity of infection or condition, the age, sex, weight or medical condition of the subject and the route of administration. In that regard, the precise amount of the compound that is to be administered may depend on the judgement of the practitioner.
• the subject can be a mammal.
• the subject is a human, a companion animal (e.g., dog, cat, ferret, hamster, gerbil, etc.), a livestock animal (e.g., cattle, pig, horse, poultry, etc.), or any other mammal in need of treatment.
• the compounds described herein may be administered over a period of hours, days, weeks, or months, depending on several factors, including the severity of the infection or condition being treated, whether a recurrence is considered likely.
• the administration may be via infusion over a period of hours, days, weeks, months, etc.
• the administration may be intermittent, e.g., once per day over a period of days, once per hour over a period of hours, or any other such schedule as deemed suitable.
• the composition of the present disclosure is administered once daily for at least one week, for example, at least once daily for at least two weeks, or once daily for at least one month or longer.
• the composition of the present disclosure is provided immediately prior, or subsequent, to exposure to an infectious agent, or upon onset of risk for a bone infection.
• the compound of the present disclosure is administered at an amount of from about 0.01 mg/kg to 1000 mg/kg of body weight.
• the compound of the present disclosure may be administered at an amount of from about 0.1 mg/kg to 900 mg/kg, or from about 0.5 mg/kg to 800 mg/kg, or from about 1 mg/kg to 750 mg/kg, or from about 1.5 mg/kg to 700 mg/kg, or from about 2 mg/kg to 600 mg/kg, or from about 2 mg/kg to 500 mg/kg, or from about 2.5 mg/kg to 450 mg/kg, or from about 3 mg/kg to 350 mg/kg, or from about 3.5 mg/kg to 250 mg/kg, or from about 4 mg/kg to 200 mg/kg, or from about 4 mg/kg to 100 mg/kg, or from about 4 mg/kg to 50 mg/kg, or from about 4 mg/kg to 20 mg/kg, or any dose bounded by these ranges.
• the compound of the present disclosure is administered at about 2 mg/kg, about 5 mg/kg, about 8 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, about 30 mg/kg, about 35 mg/kg, about 40 mg/kg, about 45 mg/kg, or about 50 mg/kg, or within any range bounded by the foregoing dosage amounts.
• the antibiotic compound is administered systemically.
• the targeting conferred by the bone-binding moiety may reduce the dosing that is required to provide an effective amount of the antibiotic (e.g., CSA) at the site of infection compared to the dosing that would be required in the absence of a bone-binding moiety. It will also be understood that higher doses of the bone-binding compound may be tolerated by a patient compared to the doses that may be tolerated of an antibiotic compound lacking bone-binding capabilities, as the bone-binding antibiotic compound is directed to the tissue of interest rather than travelling non-specifically to other locations in the body.
• the antibiotic e.g., CSA
• the present disclosure provides an oral dose of from about 0.01 mg to 4000 mg of the active ingredient, such as from about 0.05 mg to 3500 mg, or from about 0.1 mg to 3000 mg, from about 0.5 mg to 2500 mg, from about 0.75 mg to 2000 mg, or from about 1 mg to 1750 mg, from about 1.25 mg to 1500 mg, or from about 1.5 mg to 1250 mg, or from about 2 mg to 1000 mg, or from about 5 mg to 900 mg, from about 7.5 mg to 800 mg, or from about 10 mg to 700 mg, or from about 15 mg to 600 mg, or from about 20 mg to 550 mg, or from about 25 mg to 500 mg, or from about 30 mg to 500 mg, or from about 35 mg to 450 mg, or from about 40 mg to 450 mg, or from about 45 mg to 450 mg, or from about 50 mg to 400 mg.
• the active ingredient such as from about 0.05 mg to 3500 mg, or from about 0.1 mg to 3000 mg, from about 0.5 mg to 2500 mg, from about 0.75 mg to 2000 mg, or
• Suitable routes may, for example, include oral, rectal, transmucosal, or intestinal administration; parenteral delivery, including intramuscular, subcutaneous, transcutaneous, intradermal, or intramedullary delivery (e.g. , injection), as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, pulmonary, transdermal, or intraocular delivery (e.g., injection).
• a release system can include a matrix of a biodegradable material or a material which releases the incorporated components by diffusion.
• the components can be homogeneously or heterogeneously distributed within the release system.
• release systems may be useful, however, the choice of the appropriate system will depend upon rate of release required by a particular application. Both non-degradable and degradable release systems can be used.
• Suitable release systems include polymers and polymeric matrices, non-polymeric matrices, or inorganic and organic excipients and diluents such as, but not limited to, calcium carbonate and sugar (for example, trehalose).
• the release system material can be selected so that components having different molecular weights are released by diffusion or through degradation of the material.
• Representative synthetic, biodegradable polymers include, for example: polyamides such as poly(amino acids) and poly(peptides); polyesters such as poly(lactic acid), poly(glycolic acid), poly(lactic-co-glycolic acid), and poly(caprolactone); poly(anhydrides); polyorthoesters; polycarbonates; and chemical derivatives thereof (substitutions, additions of chemical groups, for example, alkyl, alkylene, hydroxylations, oxidations, and other modifications routinely made by those skilled in the art), copolymers and mixtures thereof.
• Representative synthetic, non-degradable polymers include, for example: poly ethers such as poly(ethylene oxide), poly(ethylene glycol), and poly(tetramethylene oxide); vinyl polymers-polyacrylates and polymethacrylates such as methyl, ethyl, other alkyl, hydroxyethyl methacrylate, acrylic and methacrylic acids, and others such as poly(vinyl alcohol), poly(vinyl pyrolidone), and poly(vinyl acetate); poly (urethanes); cellulose and its derivatives, such as alkyl, hydroxyalkyl, ethers, esters, nitrocellulose, and various cellulose acetates; polysiloxanes; and any chemical derivatives thereof (substitutions, additions of chemical groups, for example, alkyl, alkylene, hydroxylations, oxidations, and other modifications routinely made by those skilled in the art), copolymers and mixtures thereof.
• antibiotic agents may include, for example, ciprofloxacin, gemcitabine, tryptophan, paclitaxel, cytarabine, rifalazil, norfloxacin, enoxacin, gatifloxacin, moxifloxacin, a fluoroquinolone ester, a benzoxazinorifamycine, aminoglycoside, polyene, nitroimidazole, rifamycin, bacitracin, a beta-lactam, cephalosporin, chloramphenicol, a gly copeptide, a macrolide, a lincosamide, penicillin, a quinolones, rifampicin, tetracycline, trimethoprim a sulfonamide, amoxicillin, augmentin, amoxicillin, ampicillin, azlocillin, flucloxacillin
• the compounds of the present disclosure may be administered in combination with additional compounds that are useful for promoting bone formation or decreasing bone resorption.
• suitable compounds may include risedronate (Actonel), Ibandronate (Boniva), or zoledronic acid (Reclast or Aclasta).
• the other compound may be a corticosteroid, e.g., prednisone or cortisone.
• the other compound may be denosumab (Prolia).
• the other compound may be strontium ranelate (Protos).
• the other compound may be a selective estrogen receptor modulator (SERMS), such as raloxifene (Evista).
• SERMS selective estrogen receptor modulator
• the other compound may be a drug used in hormone replacement therapy (HRT), such as estrogen or progesterone.
• HRT hormone replacement therapy
• the other compound may be teriparatide (Forteo).
• the other compound may be a non-steroidal anti inflammatory agent or analgesic.
• a suitable non-steroidal anti-inflammatory agent may be ibuprofen, naproxen, aspirin, or a COX-1 and/or COX-2 inhibitor selected from ketoprofen, indomethacin (Indocin or Tivorbex), and fenoprofen (Nalfon).
• the compound of the present disclosure may be administered to a subject in association with a scaffold.
• the scaffold material may be as described in U.S. Patent Nos. 5,681,872; 5,914,356; 5,939,039; 6,325,987; 6,383,519; 6,521,246; 6,736,799; 6,800,245; 6,969,501; 6,991,803; 7,052,517; 7,189,263; 7,534,451; 8,303,967; 8,460,686; or
• scaffold materials may include VITOSS®, CORTOSS®, biopolymers, bone, decellularized bone, extracellular matrix or components thereof, fibronectin, laminin collagen, chitosan, alginate, calcium phosphate, calcium sulfate, poly(alpha-hydroxy acids) such as poly(lactic-co-glycolic acid) and polyglycolic acid, CUPE polymer, polyethylene glycol, or any combinations thereof.
• the scaffold material may be porous.
• the scaffold material may be a natural material, synthetic material, or a combination thereof.
• the scaffold material may be biocompatible, nontoxic and/or non-inflammatory.
• the scaffold material may support cell attachment, cell proliferation, extracellular and/or bone matrix production, and/or cell conversion.
• the scaffold material may be biodegradable.
• the scaffold material may be sterilized.
• Other scaffold materials and attributes will be appreciated by those of skill in the art.
• the compounds of the present disclosure may be administered by a variety of routes.
• the compound is administered systemically such as by direct delivery into the bloodstream of a subject.
• the compound is delivered parenterally.
• routes of parenteral administration include, but are not limited to, intravascular, intracapsular, intraorbital, intracardiac, intradermal, transtacheal, intraperitoneal, intraventricular, intracerebroventricular, intrathecal, subcutaneous, subcuticular, subcapsular, subarachnoid, intraspinal, epidural, intrastemal, intracranial, intramuscular, intraarticular, intra-arterial, intranodal, pulmonary, intranasal, transdermal, and intravenous.
• the compound is administered intraperitoneally or intravenously.
• the compounds may be formulated using pharmaceutically acceptable carriers well known in the art into dosage forms suitable for oral administration such as tablets, pills, capsules, liquids, gels, syrups, slurries, suspensions, and the like.
• suitable carriers may be selected from malt, gelatin, talc, calcium sulphate, vegetable oils, synthetic oils, polyols, alginic acid, phosphate buffered solutions, emulsifiers, isotonic saline, and pyrogen-free water.
• the compound is administered intranasally or by inhalation (for example, in the form of an aerosol spray presentation from a pressurized container, pump, spray or nebulizer with the use of a suitable propellant, such as dichlorodifluoromethane, trichlorofluoro-methane, dichlorotetra- fluoro-ethane, a hydrofluoroalkane such as 1,1,1,2-tetrafluoroethane (HFA 134A3 or l,l,l,2,3,3,3-heptafluoropropane (HFA227EA3), carbon dioxide, or other suitable gas) or as solid micronized powder delivered with dry powder inhaler
• a suitable propellant such as dichlorodifluoromethane, trichlorofluoro-methane, dichlorotetra- fluoro-ethane, a hydrofluoroalkane such as 1,1,1,2-tetrafluoroethane (HFA
• Bone-Binding Antibiotic-1 (BBA-1) was synthesized in a two-step reaction.
• a NHS-PEG-COOH linker (1 kDa, 33 mg) was dissolved in 1 mL of Milli-Q water.
• alendronate (ALN) sodium (10.5 mg) was weighed and dissolved in 1 mL of Milli-Q water.
• Alendronate solution was added dropwise into the solution of NHS-PEG- COOH under continuous stirring. pH of the reaction was monitored and adjusted to pH 7 using NaOH solution. The reaction mixture was stirred overnight at room temperature.
• BBA-1 has a molecular weight of about 2070.12 g/mole and its structure is:
• n is about 16.
• BBA-1 was characterized using high performance liquid chromatography (HPLC).
• HPLC high performance liquid chromatography
• a broad peak at 8.2 min and 9.3 min was attributed to the possible conjugation of ALN-PEG-COOH linker to the two primary amino groups of CSA-90.
• a peak at 7.5 min was attributed to unreacted linker, while a peak at 2.5 min was due to the EDC.HLC carbodiimide.
• Corresponding elution and identification of pure NHS- PEG-COOH linker at time 5 mins and CSA-90 at 6.8 min using similar method is shown in Figure 1-B and Figure 1-C, respectively.
• Figure 2 shows FT-IR spectrums for alendronate (Figure 2-A), CSA-90 (Figure 2- B), NHS-PEG-COOH linker (Figure 2-C) and BBA-1 ( Figure 2-D), respectively.
• the existence of CSA-90 in the spectrum of BBA-1 ( Figure 2-D) was confirmed by the identification of peaks at 2867 cm 1 and 2925 cm 1 . In all spectra, the peaks around 1550- 1560 cm 1 and above 3300 cm 1 correspond to N-H bending and stretching vibrations, respectively ( Figure 2-A to 2-D).
• alendronate was conjugated to the NHS terminus of the NHS-PEG-COOH linker.
• CSA-90 was conjugated to the COOH terminus of the linker using water soluble l-ethyl-3-(3-dimethylaminopropyl)- carbodiimide (EDC.HCL).
• EDC.HCL water soluble l-ethyl-3-(3-dimethylaminopropyl)- carbodiimide
• Figure 3-A shows the 'H NMR spectrums for BBA- 1, NHS-PEG-COOH, CSA-90, and alendronate sodium. The success of alendronate conjugation was confirmed by 'H NMR and 31 P NMR spectroscopy.
• a modified Kirby -Bauer disc diffusion assay was performed to test the bactericidal activity of BBA-1 against common species of bacteria causing bone infection.
• MRSA methicillin-resistant Staphylococcus aureus
• minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of BBA- 1 was determined by standardized broth microdilution method. Briefly, 1 x 10 5 CFU/mL of S .aureus and MRSA inoculum was incubated with serial concentrations of BBA-1 and CSA-90. The optical density/absorbance was recorded in triplicate at 595 nm after 24 hours of incubation at 37 °C.
• BBA-1 -treated discs significantly inhibited bacterial growth across all experiments.
• the antibacterial activity of BBA-1 at an equimolar concentration of CSA-90 and gentamicin was comparable against both strains of bacteria.
• the MIC and MBC concentration of BBA-1 against S. aureus and MRSA were 1.5 pg/mL and 3 pg/mL, respectively ( Figure 4-C).
• HA binding affinity of BBA-1 and CSA-90 was determined semi-quantitatively by incubating each compound with and without HA in milli-Q water.
• BBA-1 (2.42 mg) was weighed and dissolved in 4 mL of milli-Q water.
• HA (20 mg) was weighed and suspended in 2 mL of BBA-1 solution, and the suspension was incubated at room temperature.
• 100 pL of supernatant was withdrawn at predetermined time points and analyzed by HPLC.
• bone binding ability of pure CSA- 90 was tested by incubating CSA-90 with and without HA at similar concentrations.
• CSA-90 has been reported to have pro-osteogenic activity and potentiate recombinant human bone morphogenetic protein-2 (rhBMP-2) in cultured cells (Schindeler et al, JBone Joint Surg Am. 97(4):302-9 (2015)). This may lead to additional benefits beyond antimicrobial protection, particularly in the context of the repair of bony injuries, bone defects, or orthopedic implant osseointegration.
• rhBMP-2 human bone morphogenetic protein-2
• MC3T3- El cells were differentiated in osteogenic media with either 5 mM CSA-90 or BBA-1, with or without 50 ng/ml rhBMP-2.
• a p-nitrophenyl phosphate assay was performed for alkaline phosphatase activity (Sigma- Aldrich) and normalized to day-3 cells grown in osteogenic media and a-MEM media. Assays were performed in triplicate with two independent repeats.
• parental compound alendronate is a bisphosphonate that impacts on bone resorption by functionally affecting the mevalonate pathway. It was hypothesized by conjugation by the sidechain amine group, the bone binding affinity would be retained, but the anti-resorptive activity would be abrogated.
• the model featured a drill hole made in the tibia of C57BL6 mice that was inoculated with live Staphylococcus aureus (strain ATCC 12600) at the time of surgery (1E5 CFU).
• mice that did not receive infection did not test positive on swab assays of the soft tissue or pin.
• Numbers of infected samples and levels of infection as measured by optical density (OD595) when culturing for local infection were reduced in the group treated with BBA-1 compared to the infected controls that did not receive BBA-1.

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Chemical & Material Sciences (AREA)
• General Health & Medical Sciences (AREA)
• Medicinal Chemistry (AREA)
• Pharmacology & Pharmacy (AREA)
• Animal Behavior & Ethology (AREA)
• Public Health (AREA)
• Veterinary Medicine (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Epidemiology (AREA)
• Organic Chemistry (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• General Chemical & Material Sciences (AREA)
• Physical Education & Sports Medicine (AREA)
• Orthopedic Medicine & Surgery (AREA)
• Biophysics (AREA)
• Molecular Biology (AREA)
• Rheumatology (AREA)
• Communicable Diseases (AREA)
• Oncology (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Medicinal Preparation (AREA)
• Steroid Compounds (AREA)

Priority Applications (7)

Applications Claiming Priority (2)

Publications (1)

Family

ID=74194302

Family Applications (1)

Country Status (8)

Cited By (1)

Families Citing this family (2)

Citations (4)

Family Cites Families (6)

• 2020
  • 2020-07-24 CN CN202080066011.3A patent/CN114585316A/zh active Pending
  • 2020-07-24 CA CA3145403A patent/CA3145403A1/en active Pending
  • 2020-07-24 AU AU2020319089A patent/AU2020319089A1/en not_active Abandoned
  • 2020-07-24 EP EP20844850.6A patent/EP4003201A4/en active Pending
  • 2020-07-24 KR KR1020227006236A patent/KR20220041152A/ko not_active Ceased
  • 2020-07-24 WO PCT/US2020/043563 patent/WO2021016583A1/en not_active Ceased
  • 2020-07-24 US US17/629,058 patent/US20220273673A1/en active Pending
  • 2020-07-24 JP JP2022504548A patent/JP7580815B2/ja active Active

Patent Citations (4)

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events