US20130224232A1 - Cyclic tetrapeptides and therapeutic applications thereof - Google Patents

Cyclic tetrapeptides and therapeutic applications thereof Download PDF

Info

Publication number
US20130224232A1
US20130224232A1 US13/579,933 US201113579933A US2013224232A1 US 20130224232 A1 US20130224232 A1 US 20130224232A1 US 201113579933 A US201113579933 A US 201113579933A US 2013224232 A1 US2013224232 A1 US 2013224232A1
Authority
US
United States
Prior art keywords
compound
formula
taken together
attached
alkyl
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US13/579,933
Other languages
English (en)
Inventor
Janusz ZABROCKI
Michal Zimecki
Andrzij Kaszuba
Krzysztof KACZMAREK
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
LODZ UNIVERSITY OF TECHNOLOGY
Peptaderm Sp Z Oo
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to US13/593,716 priority Critical patent/US9382292B2/en
Publication of US20130224232A1 publication Critical patent/US20130224232A1/en
Assigned to LODZ UNIVERSITY OF TECHNOLOGY, PEPTADERM SP. Z O.O. reassignment LODZ UNIVERSITY OF TECHNOLOGY ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: KACZMAREK, KRZYSZTOF, KASZUBA, ANDRZEJ, ZIMECKI, MICHAL, ZABROCKI, Janusz
Abandoned legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/12—Cyclic peptides with only normal peptide bonds in the ring
    • C07K5/126—Tetrapeptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/12—Cyclic peptides, e.g. bacitracins; Polymyxins; Gramicidins S, C; Tyrocidins A, B or C
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/08—Antiseborrheics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains three hetero rings
    • C07D487/14—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0202—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-X-X-C(=0)-, X being an optionally substituted carbon atom or a heteroatom, e.g. beta-amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/12—Cyclic peptides with only normal peptide bonds in the ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/50—Cyclic peptides containing at least one abnormal peptide link
    • C07K7/54—Cyclic peptides containing at least one abnormal peptide link with at least one abnormal peptide link in the ring
    • C07K7/56—Cyclic peptides containing at least one abnormal peptide link with at least one abnormal peptide link in the ring the cyclisation not occurring through 2,4-diamino-butanoic acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/64—Cyclic peptides containing only normal peptide links

Definitions

  • Immunosuppressive drugs are commonly used in transplantation and in treatment of autoimmune diseases. Production of these drugs is expensive, and the most frequently used of these drugs, namely cyclosporine A, tacrolimus and rapamycin, exhibit undesirable side-effects.
  • Cyclolinopeptide A (CLA), a very hydrophobic cyclic nonapeptide, was first isolated from linen seeds in 1959. CLA is strongly immunosuppressive, with a potency comparable to that of cyclosporine A (CsA). The mechanism of action of CLA was shown to be similar to that of CsA, i.e. CLA formed a complex with cyclophilin A causing inactivation of calcineurin, albeit at much lower affinity (Gaymes et al., Febs Lett, 1997, 418, 224-227).
  • CLA inhibited both humoral and cellular immune response and graft-versus-host reaction; prolonged survival of allogeneic skin grafts; tempered post-adjuvant polyarthritis in rats and hemolytic anemia of New Zealand Black mice; and, similarly to CsA, inhibited IL1 and IL-2 production.
  • CsA inhibited IL1 and IL-2 production.
  • the high hydrophobicity of CLA presents an obstacle for the potential application of the compound in therapy.
  • Linear CLA analogues containing alanine residue in successive positions of the peptide chain were found to be immunosuppressive (Wieczorek et al., Arch Immunol Ther Exp, 1992, 40, 213-216). It was also found that the activity of linear CLA analogues gradually decreased with shortening of the peptide chain from the N-terminus, at the same time showing an increase of activity for C-terminal tetra- and tripeptides (Siemion et al., Arch Immunol Ther Exp, 1994, 42, 459-465). The introduction of a single, hydrophilic threonine residue into the CLA molecule did not result in improved solubility in water.
  • Synthetic CLA analogues in which leucine residues in position 5 and/or 8 were replaced with their hydroxymethyl analogue displayed a four-fold increase in solubility in water in comparison to CLA, but also showed a 25% diminution in biological activity compared to native CLA (Zubrzak et al., Biopolymers (Peptide Science), 2005, 80, 347-356).
  • CLA analogues A series of nine CLA analogues was obtained by replacement of CLA proline residues with ⁇ 2 -isoproline and ⁇ 3 -homoproline. In comparison to CsA, these CLA analogues displayed strong inhibitory properties in the cellular immune response. The majority of these analogs were practically devoid of cell toxicity (Katarzy ⁇ ska et al., J Pept Sci, 2009, 14, 1283-1294).
  • k, m, n and p are each independently 0, 1 or 2;
  • R and R′ are each independently selected from H and C 1-3 alkyl, or, when taken together, R and R′ are —CR 1 R 1′ —X—CH 2 —, wherein CR 1 R 1′ is attached to the backbone nitrogen, R 1 and R 1′ are each independently selected from H and C 1-3 alkyl, and X is selected from —CH 2 —, —CH 2 CH 2 —, —CH(OH)—, —O—, —S— and —NH—;
  • R′′ and R′′′ are each independently selected from H and C 1-3 alkyl, or, when taken together, R′′ and R′′′ are —CR 2 R 2′ —X′—CH 2 —, wherein CR 2 R 2′ is attached to the backbone nitrogen, R 2 and R 2′ are each independently selected from H and C 1-3 alkyl, and X′ is selected from —CH 2 —, —CH 2 CH 2 —, —CH(OH)—, —O—, —S— and —NH—; and
  • R 3 and R 4 are each independently selected from aryl, substituted aryl, heteroaryl and substituted heteroaryl;
  • At least one of R 3 and R 4 is phenyl. In some embodiments, at least one of R 3 and R 4 is 4-hydroxyphenyl. In some embodiments, at least one of R 3 and R 4 is 4-t-butoxyphenyl. In some embodiments, at least one of R 3 and R 4 is 2-indolyl. In some embodiments, R 3 and R 4 are both phenyl. In some embodiments, one of R 3 and R 4 is phenyl and the other of R 3 and R 4 is 4-hydroxyphenyl. In some embodiments, one of R 3 and R 4 is phenyl and the other of R 3 and R 4 is 4-t-butoxyphenyl.
  • one of R 3 and R 4 is phenyl and the other of R 3 and R 4 is 2-indolyl.
  • the carbon to which —CH 2 —R 3 is attached has absolute (R)-stereochemistry.
  • the carbon to which —CH 2 —R 3 is attached has absolute (S)-stereochemistry.
  • the carbon to which —CH 2 —R 4 is attached has absolute (R)-stereochemistry.
  • the carbon to which —CH 2 —R 4 is attached has absolute (S)-stereochemistry.
  • one of k, m, n and p is 1, and the remainder of k, m, n and p are 0.
  • two of k, m, n and p are 1, and the remainder of k, m, n and p are 0.
  • at least one of k and m is not 0.
  • at least one of n and p is not 0.
  • at least one of k and m is not 0 and at least one of n and p is not 0.
  • both k and n are 0.
  • both k and n are 0, one of m and p is 0, and the other of m and p is 1.
  • both k and n are 0 and both m and p are 1.
  • all four amino acids are L-amino acids.
  • three of the amino acids are L-amino acids and one of the amino acids is a D-amino acid. In some embodiments, two of the amino acids are L-amino acids and two of the amino acids are D-amino acids. In some embodiments, one of the amino acids is an L-amino acid and three of the amino acids are D-amino acids. In some embodiments, all four amino acids are D-amino acids.
  • R and R′ are taken together to form —(CH 2 ) 3 —, i.e. R and R′ are taken together to form —CR 1 R 1′ —X—CH 2 — wherein R 1 and R 1′ are both H and X is CH 2 .
  • R′′ and R′′′ are taken together to form —(CH 2 ) 3 —, i.e. R′′ and R′′′ are taken together to form —CR 2 R 2′ —X′—CH 2 — wherein R 2 and R 2′ are both H and X′ is CH 2 .
  • R and R′ are taken together to form —(CH 2 ) 4 —, i.e.
  • R and R′ are taken together to form —CR 1 R 1 —X—CH 2 — wherein R 1 and R 1′ are both H and X is (CH 2 ) 2 .
  • R′′ and R′′′ are taken together to form —(CH 2 ) 4 —, i.e. R′′ and R′′′ are taken together to form —CR 2 R 2′ —X—CH 2 — wherein R 2 and R 2′ are both H and X is (CH 2 ) 2 .
  • R and R′ are taken together to form —CH 2 —CH(OH)—CH 2 —, i.e.
  • R and R′ are taken together to form —CR 1 R 1′ —X—CH 2 — wherein R 1 and R 1′ are both H and X is CH(OH).
  • R′′ and R′′′ are taken together to form —CH 2 —CH(OH)—CH 2 —, i.e. R′′ and R′′′ are taken together to form —CR 2 R 2′ —X′—CH 2 — wherein R 2 and R 2′ are both H and X′ is CH(OH).
  • the carbon at which R′ is attached has absolute (S)-stereochemistry.
  • the carbon at which R′ is attached has absolute (R)-stereochemistry. In some embodiments in which R′′ and R′′′ are taken together, the carbon at which R′′′ is attached has absolute (S)-stereochemistry. In some embodiments in which R′′ and R′′′ are taken together, the carbon at which R′′′ is attached has absolute (R)-stereochemistry.
  • the compound is selected from the group consisting of:
  • the compound is a compound of formula I-1. In some embodiments, the compound is a compound of formula I-2. In some embodiments, the compound is a compound of formula I-3. In some embodiments, the compound is a compound of formula I-4. In some embodiments, the compound is a compound of formula I-5. In some embodiments, the compound is a compound of formula I-6. In some embodiments, the compound is a compound of formula I-7. In some embodiments, the compound is a compound of formula I-8.
  • the compound is selected from the group consisting of:
  • the compound is a compound of formula I-A. In some embodiments, the compound is a compound of formula I-B. In some embodiments, the compound is a compound of formula I-C. In some embodiments, the compound is a compound of formula I-D. In some embodiments, the compound is a compound of formula I-E. In some embodiments, the compound is a compound of formula I-F. In some embodiments, the compound is a compound of formula I-G. In some embodiments, the compound is a compound of formula I-H. In some embodiments, the compound is a compound of formula I-I. In some embodiments, the compound is a compound of formula I-J. In some embodiments, the compound is a compound of formula I-K.
  • the compound is a compound of formula I-L. In some embodiments, the compound is a compound of formula I-M. In some embodiments, the compound is a compound of formula I-N. In some embodiments, the compound is a compound of formula I-O. In some embodiments, the compound is a compound of formula I-P.
  • one or more amino groups in the compound of Formula I are in protected form.
  • a pharma-ceutical composition comprising a compound of formula I and a pharmaceutically acceptable carrier, excipient or diluent therefor.
  • the compound is a compound of formula I-1.
  • the compound is a compound of formula I-2.
  • the compound is a compound of formula I-3.
  • the compound is a compound of formula I-4.
  • the compound is a compound of formula I-5.
  • the compound is a compound of formula I-6.
  • the compound is a compound of formula I-7.
  • the compound is a compound of formula I-8.
  • the compound is a compound of formula I-A. In some embodiments, the compound is a compound of formula I-B. In some embodi-ments, the compound is a compound of formula I-C. In some embodiments, the compound is a compound of formula I-D. In some embodiments, the compound is a compound of formula I-E. In some embodiments, the compound is a compound of formula I-F. In some embodi-ments, the compound is a compound of formula I-G. In some embodiments, the compound is a compound of formula I-H. In some embodiments, the compound is a compound of formula I-I. In some embodiments, the compound is a compound of formula I-J. In some embodiments, the compound is a compound of formula I-K.
  • the compound is a compound of formula I-L. In some embodiments, the compound is a compound of formula I-M. In some embodiments, the compound is a compound of formula I-N. In some embodiments, the compound is a compound of formula I-O. In some embodiments, the compound is a compound of formula I-P.
  • a method of suppressing immune response in a patient comprising administering to a patient in need thereof an efficacious amount of a compound of formula I.
  • the immune response which is suppressed is inflammation.
  • the immune response which is suppressed is transplant rejection.
  • the compound is a compound of formula I-1.
  • the compound is a compound of formula I-2.
  • the compound is a compound of formula I-3.
  • the compound is a compound of formula I-4.
  • the compound is a compound of formula I-5.
  • the compound is a compound of formula I-6.
  • the compound is a compound of formula I-7. In some embodiments, the compound is a compound of formula I-8. In some embodiments, the compound is a compound of formula I-A. In some embodiments, the compound is a compound of formula I-B. In some embodiments, the compound is a compound of formula I-C. In some embodiments, the compound is a compound of formula I-D. In some embodiments, the compound is a compound of formula I-E. In some embodiments, the compound is a compound of formula I-F. In some embodiments, the compound is a compound of formula I-G. In some embodiments, the compound is a compound of formula I-H. In some embodiments, the compound is a compound of formula I-I.
  • the compound is a compound of formula I-J. In some embodiments, the compound is a compound of formula I-K. In some embodiments, the compound is a compound of formula I-L. In some embodiments, the compound is a compound of formula I-M. In some embodiments, the compound is a compound of formula I-N. In some embodiments, the compound is a compound of formula I-O. In some embodiments, the compound is a compound of formula I-P.
  • a method of treating or preventing an immune-mediated disease or condition in a patient comprising administering to a patient in need thereof an efficacious amount of a compound of formula I.
  • a method for lowering the toxicity profile of a second drug comprising administering a compound of formula I in conjunction with the second drug.
  • the immune-mediated disease or condition is selected from the group consisting of auto-immune diseases, inflam-mation processes, transplant rejection, and allergic reactions.
  • the immune-mediated disease or condition is selected from Psoriasis, lichen planus and other papulosquamous disorders.
  • the immune-mediated disease or condition is selected from eczema and dermatitis.
  • the eczemea or dermatitis is selected from eczema, atopic eczema, seborrheic dermatitis, and pompholyx.
  • the immune-mediated disease or condition is a skin reaction to sunlight.
  • the immune-mediated disease or condition selected from non-specific skin irritation and insect bite.
  • the immune-mediated disease or condition is a urticaria.
  • the immune-mediated disease or condition is selected from the group consisting of a primary skin tumor (e.g.
  • the compound of formula I is administered in conjunction with a chemotherapeutic drug in order to reduce the toxic effects of the chemotherapeutic drug.
  • the compound is a compound of formula I-1.
  • the compound is a compound of formula I-2.
  • the compound is a compound of formula I-3.
  • the compound is a compound of formula I-4. In some embodiments, the compound is a compound of formula I-5. In some embodiments, the compound is a compound of formula I-6. In some embodiments, the compound is a compound of formula I-7. In some embodiments, the compound is a compound of formula I-8. In some embodiments, the compound is a compound of formula I-A. In some embodiments, the compound is a compound of formula I-B. In some embodiments, the compound is a compound of formula I-C. In some embodiments, the compound is a compound of formula I-D. In some embodiments, the compound is a compound of formula I-E. In some embodiments, the compound is a compound of formula I-F.
  • the compound is a compound of formula I-G. In some embodiments, the compound is a compound of formula I-H. In some embodiments, the compound is a compound of formula I-I. In some embodiments, the compound is a compound of formula I-J. In some em-bodiments, the compound is a compound of formula I-K. In some embodiments, the compound is a compound of formula I-L. In some embodiments, the compound is a compound of formula I-M. In some embodiments, the compound is a compound of formula I-N. In some embodiments, the compound is a compound of formula I-O. In some embodiments, the compound is a compound of formula I-P.
  • kits comprising a compound of formula I and instructions for using the compound to (a) suppress an immune response in a patient, (b) treat or prevent an immune-mediated disease or condition in a patient, or (c) lower the toxicity profile of a second drug.
  • the immune response is inflammation.
  • the immune response is transplant rejection.
  • the immune-mediated disease or condition is selected from the group consisting of auto-immune diseases, inflammation processes, transplant rejection, allergic reactions.
  • the immune-mediated disease or condition is selected from Psoriasis, lichen planus and other papulosquamous disorders.
  • the immune-mediated disease or condition is selected from eczema and dermatitis.
  • the eczemea or dermatitis is selected from eczema, atopic eczema, sebor-rheic dermatitis, and pompholyx.
  • the immune-mediated disease or con-dition is a skin reaction to sunlight.
  • the immune-mediated disease or condition selected from non-specific skin irritation and insect bite.
  • the immune-mediated disease or condition is a urticaria.
  • the immune-mediated disease or condition is selected from the group consisting of a primary skin tumor (e.g.
  • the instructions instruct administering the compound of formula I in conjunction with a chemotherapeutic drug in order to reduce the toxic effects of the chemotherapeutic drug.
  • the compound is a compound of formula I-1.
  • the compound is a compound of formula I-2.
  • the compound is a compound of formula I-3.
  • the compound is a compound of formula I-4. In some embodiments, the compound is a compound of formula I-5. In some embodiments, the compound is a compound of formula I-6. In some embodiments, the compound is a compound of formula I-7. In some embodiments, the compound is a compound of formula I-8. In some embodiments, the compound is a compound of formula I-A. In some embodiments, the compound is a compound of formula I-B. In some embodiments, the compound is a compound of formula I-C. In some embodiments, the compound is a compound of formula I-D. In some embodiments, the compound is a compound of formula I-E. In some embodiments, the compound is a compound of formula I-F.
  • the compound is a compound of formula I-G. In some embodiments, the compound is a compound of formula I-H. In some embodiments, the compound is a compound of formula I-I. In some embodiments, the compound is a compound of formula I-J. In some embodiments, the compound is a compound of formula I-K. In some embodiments, the compound is a compound of formula I-L. In some embodiments, the compound is a compound of formula I-M. In some embodiments, the compound is a compound of formula I-N. In some embodiments, the compound is a compound of formula I-O. In some embodiments, the compound is a compound of formula I-P.
  • a method for making a compound of formula I comprising cyclizing a compound having the formula II-1, II-2, II-3 or II-4, wherein R, R′, R′′, R′′′, R 3 , R 4 , k, m, n and p are as defined in formula I (collectively referred to hereinafter as compounds of formula II):
  • the method further comprises synthesizing the compound of formula II-1, II-2, II-3 or II-4.
  • the compound of formula II-1, II-2, II-3 or II-4 is formed by solid-phase synthesis.
  • compounds of formulae II-1, II-2, II-3 and II-4 per se, as well as protected versions of these compounds (e.g. in which one or more amino groups, such as the N-terminal amino group or a side chain amino group, are protected, e.g. by tert-butoxycarbonyl) and these compounds, in protected or unprotected form, when bound to a solid-phase resin.
  • protected versions of these compounds e.g. in which one or more amino groups, such as the N-terminal amino group or a side chain amino group, are protected, e.g. by tert-butoxycarbonyl
  • the compound of formula II is selected from the group consisting of:
  • the compound is a compound of formula II-4-a. In some embodiments, the compound is a compound of formula II-1-a. In some embodiments, the compound is a compound of formula II-2-a. In some embodiments, the compound is a compound of formula II-3-a. In some embodiments, the compound is a compound of formula II-4-b. In some embodiments, the compound is a compound of formula II-1-b. In some embodiments, the compound is a compound of formula II-2-b. In some embodiments, the compound is a compound of formula II-3-b. In some embodiments, the compound is a compound of formula II-4-c. In some embodiments, the compound is a compound of formula II-1-c.
  • the compound is a compound of formula II-2-c. In some embodiments, the compound is a compound of formula II-3-c. In some embodiments, the compound is a compound of formula II-4-d. In some embodiments, the compound is a compound of formula II-1-d. In some embodiments, the compound is a compound of formula II-2-d. In some embodiments, the compound is a compound of formula II-3-d. In some embodiments, the compound is a compound of formula II-4-e. In some embodiments, the compound is a compound of formula II-1-e. In some embodiments, the compound is a compound of formula II-2-e. In some embodiments, the compound is a compound of formula II-3-e.
  • the compound is a compound of formula II-4-f. In some embodiments, the compound is a compound of formula II-1-f. In some embodiments, the compound is a compound of formula II-2-f. In some embodiments, the compound is a compound of formula II-3-f. In some embodiments, the compound is a compound of formula II-4-g. In some embodiments, the compound is compound of formula II-1-g. In some embodiments, the compound is compound of formula II-2-g. In some embodiments, the compound is compound of formula II-3-g. In some embodiments, the compound is a compound of formula II-4-h. In some embodiments, the compound is compound of formula II-1-h. In some embodiments, the compound is compound of formula II-2-h. In some embodiments, the compound is compound of formula II-3-h.
  • the compound is a compound of formula II-A-1. In some embodiments, the compound is a compound of formula II-A-2. In some embodiments, the compound is a compound of formula II-A-3. In some embodiments, the compound is a compound of formula II-A-4. In some embodiments, the compound is a compound of formula II-B-1. In some embodiments, the compound is a compound of formula II-B-2. In some embodi-ments, the compound is a compound of formula II-B-3. In some embodiments, the compound is a compound of formula II-B-4. In some embodiments, the compound is a compound of formula II-C-1. In some embodiments, the compound is a compound of formula II-C-2.
  • the compound is a compound of formula II-C-3. In some embodiments, the compound is a compound of formula II-C-4. In some embodiments, the compound is a compound of formula II-D-1. In some embodiments, the compound is a compound of formula II-D-2. In some embodiments, the compound is a compound of formula II-D-3. In some embodiments, the compound is a compound of formula II-D-4. In some embodiments, the compound is a compound of formula II-E-1. In some embodiments, the compound is a compound of formula II-E-2. In some embodiments, the compound is a compound of formula II-E-3. In some embodiments, the compound is a compound of formula II-E-4.
  • the compound is a compound of formula II-F-1. In some embodiments, the compound is a compound of formula II-F-2. In some embodiments, the compound is a compound of formula II-F-3. In some embodiments, the compound is a compound of formula II-F-4. In some embodiments, the compound is a compound of formula II-G-1. In some embodiments, the compound is a compound of formula II-G-2. In some embodiments, the compound is a compound of formula II-G-3. In some embodiments, the compound is a compound of formula II-G-4. In some embodiments, the compound is a compound of formula II-H-1. In some embodiments, the compound is a compound of formula II-H-2.
  • the compound is a compound of formula II-H-3. In some embodiments, the compound is a compound of formula II-H-4. In some embodiments, the compound is a compound of formula II-J-1. In some embodiments, the compound is a compound of formula II-J-2. In some embodiments, the compound is a compound of formula II-J-3. In some embodiments, the compound is a compound of formula II-J-4. In some embodiments, the compound is a compound of formula II-K-1. In some embodiments, the compound is a compound of formula II-K-2. In some embodiments, the compound is a compound of formula II-K-3. In some embodiments, the compound is a compound of formula II-K-4.
  • the compound is a compound of formula II-L-1. In some embodiments, the compound is a compound of formula II-L-2. In some embodiments, the compound is a compound of formula II-L-3. In some embodiments, the compound is a compound of formula II-L-4. In some embodiments, the compound is a compound of formula II-M-1. In some embodiments, the compound is a compound of formula II-M-2. In some embodiments, the compound is a compound of formula II-M-3. In some embodiments, the compound is a compound of formula II-M-4. In some embodiments, the compound is a compound of formula II-N-1. In some embodiments, the compound is a compound of formula II-N-2.
  • the compound is a compound of formula II-N-3. In some embodiments, the compound is a compound of formula II-N-4. In some embodiments, the compound is a compound of formula II-O-1. In some embodiments, the compound is a compound of formula II-O-2. In some embodiments, the compound is a compound of formula II-O-3. In some embodiments, the compound is a compound of formula II-O-4. In some embodiments, the compound is a compound of formula II-P-1. In some embodiments, the compound is a compound of formula II-P-2. In some embodiments, the compound is a compound of formula II-P-3. In some embodiments, the compound is a compound of formula II-P-4.
  • compounds of formula I exhibit immunosuppressive and/or anti-inflammatory activity, while at the same time exhibiting less toxicity, than some known compounds.
  • compounds of formula I may be useful as immunosuppressive and/or anti-inflammatory agents.
  • immune-mediated refers to a disease or condition in which the body's immune system overreacts and/or attacks the body.
  • Alkyl is intended to include linear, branched, or cyclic saturated hydrocarbon structures and combinations thereof.
  • Lower alkyl refers to alkyl groups of from 1 to 6 carbon atoms. Examples of lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, s- and t-butyl and the like. Preferred alkyl groups are those of C 20 or below.
  • Cycloalkyl is a subset of alkyl and includes cyclic hydrocarbon groups of from 3 to 8 carbon atoms. Examples of cycloalkyl groups include c-propyl, c-butyl, c-pentyl, norbornyl and the like.
  • C 1 to C 20 hydrocarbon includes alkyl, cycloalkyl, polycycloalkyl, alkenyl, alkynyl, aryl and combinations thereof. Examples include benzyl, phenethyl, cyclohexylmethyl, camphoryl and naphthylethyl.
  • the term “carbocycle” is intended to include ring systems consisting entirely of carbon but of any oxidation state. Thus (C 3 -C 10 ) carbocycle refers to such systems as cyclopropane, benzene and cyclohexene; (C 8 -C 12 ) carbopolycycle refers to such systems as norbornane, decalin, indane and naphthalene.
  • Alkoxy or alkoxyl refers to groups of from 1 to 8 carbon atoms of a straight, branched, cyclic configuration and combinations thereof attached to the parent structure through an oxygen atom. Examples include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy and the like. Lower-alkoxy refers to groups containing one to four carbons.
  • Oxaalkyl refers to alkyl residues in which one or more carbons has been replaced by oxygen. Examples include methoxypropoxy, 3,6,9-trioxadecyl and the like.
  • Acyl refers to groups of from 1 to 8 carbon atoms of a straight, branched, cyclic configuration, saturated, unsaturated and aromatic and combinations thereof, attached to the parent structure through a carbonyl functionality.
  • One or more carbons in the acyl residue may be replaced by nitrogen, oxygen or sulfur as long as the point of attachment to the parent remains at the carbonyl. Examples include acetyl, benzoyl, propionyl, isobutyryl, t-butoxycarbonyl, benzyloxycarbonyl and the like.
  • Lower-acyl refers to groups containing one to four carbons.
  • Aryl means 6-membered aromatic ring; a bicyclic 9- or 10-membered aromatic ring system; or a tricyclic 13- or 14-membered aromatic ring system.
  • the aromatic 6- to 14-membered carbocyclic rings include, e.g., benzene, naphthalene, indane, tetralin, and fluorene.
  • Heteroaryl mean a 5- or 6-membered heteroaromatic ring containing 1-3 heteroatoms selected from O, N, or S; a bicyclic 9- or 10-membered heteroaromatic ring system containing 1-3 heteroatoms selected from O, N, or S; or a tricyclic 13- or 14-membered heteroaromatic ring system containing 1-3 heteroatoms selected from O, N, or S.
  • the 5- to 10-membered aromatic heterocyclic rings include, e.g., imidazole, pyridine, indole, thiophene, benzopyranone, thiazole, furan, benzimidazole, quinoline, isoquinoline, quinoxaline, pyrimidine, pyrazine, tetrazole and pyrazole.
  • Arylalkyl refers to a substituent in which an aryl residue is attached to the parent structure through alkyl. Examples are benzyl, phenethyl and the like. Heteroarylalkyl refers to a substituent in which a heteroaryl residue is attached to the parent structure through alkyl. Examples include, e.g., pyridinylmethyl, pyrimidinylethyl and the like.
  • Heterocycle means a cycloalkyl or aryl residue in which from one to three carbons is replaced by a heteroatom selected from the group consisting of N, O and S.
  • the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized.
  • heterocycles include pyrrolidine, pyrazole, pyrrole, indole, quinoline, isoquinoline, tetrahydroisoquinoline, benzofuran, benzodioxan, benzodioxole (commonly referred to as methylenedioxyphenyl, when occurring as a substituent), tetrazole, morpholine, thiazole, pyridine, pyridazine, pyrimidine, thiophene, furan, oxazole, oxazoline, isoxazole, dioxane, tetrahydrofuran and the like.
  • heteroaryl is a subset of heterocycle in which the heterocycle is aromatic.
  • heterocyclyl residues additionally include piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxo-pyrrolidinyl, 2-oxoazepinyl, azepinyl, 4-piperidinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, benzimidazolyl, thiadiazolyl, benzopyranyl, benzothiazolyl, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorph
  • Substituted alkyl, aryl, cycloalkyl, heterocyclyl etc. refer to alkyl, aryl, cycloalkyl, or heterocyclyl wherein up to three H atoms in each residue are replaced with alkyl, halogen, haloalkyl, hydroxy, loweralkoxy, carboxy, carboalkoxy (also referred to as alkoxycarbonyl), carboxamido (also referred to as alkylaminocarbonyl), cyano, carbonyl, nitro, amino, alkylamino, dialkylamino, mercapto, alkylthio, sulfoxide, sulfone, acylamino, amidino, phenyl, benzyl, heteroaryl, phenoxy, benzyloxy, or heteroaryloxy.
  • halogen means fluorine, chlorine, bromine or iodine.
  • the notation “ ⁇ 3 -Ho-” (or “beta 3-homo-”) is used herein to refer to an amino acid having an extra methylene (—CH 2 —) in the backbone between the side-chain bearing carbon atom and the terminal nitrogen atom;
  • the notation “ ⁇ 2 -Ho-” (or “beta 2-homo-”) is used herein to refer to an amino acid having an extra methylene in the backbone between the side-chain bearing carbon atom and the terminal carbon atom.
  • Embodiments of the present invention include compounds of formula I in the form of salts, in particular acid addition salts.
  • Suitable salts include those formed with both organic and inorganic acids.
  • Such acid addition salts will normally be pharmaceutically acceptable, although salts of non-pharmaceutically acceptable salts may be of utility in the preparation and purification of the compound in question.
  • preferred salts include those formed from hydrochloric, hydrobromic, sulphuric, citric, tartaric, phosphoric, lactic, pyruvic, acetic, succinic, oxalic, fumaric, maleic, oxaloacetic, methanesulphonic, ethanesulphonic, p-toluenesulphonic, benzenesulphonic and isethionic acids.
  • Salts of the compounds of formula I can be made by reacting the appropriate compound in the form of the free base with the appropriate acid.
  • the compounds of formula I in accordance with the embodiments of the invention are cyclic tetrapeptides.
  • the synthesis of these peptides may be accomplished by cyclizing corresponding linear peptides that are themselves synthesized using methodologies known in the art; see, for example, Merrifield, J. Am. Chem. Soc., 85:2149 (1964); Houghten, Proc. Natl. Acad. Sci. USA, 82:5132 (1985); Kelley & Winkler in Genetic Engineering Principles and Methods , Setlow, J. K, ed., Plenum Press, N.Y., vol. 12, pp 1-19 (1990); Stewart & Young, Solid Phase Peptide Synthesis , Pierce Chemical Co., Rockford, Ill.
  • Such synthesis may be accomplish via liquid-phase or solid-phase synthesis, or by a combination of both, as is known in the art.
  • Liquid phase methods (often referred to as solution phase methods) of synthesis carry out all reactions in a homogeneous phase. Successive amino acids are coupled in solution until the desired peptide material is formed. During synthesis, successive intermediate peptides are purified by precipitation and/or washes.
  • SPPS solid phase peptide synthesis
  • a first amino acid or peptide group is bound to an insoluble support, such as a resin.
  • Successive amino acids or peptide groups are added to the first amino acid or peptide group until the peptide material of interest is formed.
  • the product of solid phase synthesis is thus a peptide bound to an insoluble support.
  • Peptides synthesized via SPPS techniques are then cleaved from the resin, and the cleaved peptide is isolated.
  • solid phase synthesis begins at the carboxy terminus of the putative peptide by coupling a protected amino acid to an inert solid support.
  • the inert solid support can be any macromolecule capable of serving as an anchor for the C-terminus of the initial amino acid.
  • the macromolecular support is a cross-linked polymeric resin (e.g. a polyamide or polystyrene resin) as shown in FIGS. 1-1 and 1-2, on pages 2 and 4 of Stewart & Young, supra.
  • the C-terminal amino acid is coupled to a polystyrene resin to form a benzyl ester.
  • a macromolecular support is selected such that the peptide anchor link is stable under the conditions used to deprotect the ⁇ -amino group of the blocked amino acids in peptide synthesis. If a base-labile ⁇ -protecting group is used, then it is desirable to use an acid-labile link between the peptide and the solid support.
  • an acid-labile ether resin is effective for base-labile Fmoc-amino acid peptide synthesis as described on page 16 of Stewart & Young, supra.
  • a peptide anchor link and ⁇ -protecting group that are differentially labile to acidolysis can be used.
  • an aminomethyl resin such as the phenylacetamidomethyl (Pam) resin works well in conjunction with Boc-amino acid peptide synthesis as described on pages 11-12 of Stewart & Young, supra. Guiller et al., Chem. Rev. 2000, 100, 2091-2157, reviewed linkers and cleavage strategies in solid-phase organic synthesis and combinatorial chemistry, including peptide synthesis.
  • the ⁇ -amino protecting group of the initial amino acid is removed with, for example, trifluoroacetic acid (TFA) in methylene chloride and neutralizing in, for example, triethylamine (TEA).
  • TFA trifluoroacetic acid
  • TAA triethylamine
  • the next ⁇ -amino and sidechain protected amino acid in the synthesis is added.
  • the remaining ⁇ -amino and, if necessary, side chain protected amino acids are then coupled sequentially in the desired order by condensation to obtain an intermediate compound connected to the solid support.
  • some amino acids may be coupled to one another to form a fragment of the desired peptide followed by addition of the peptide fragment to the growing solid phase peptide chain
  • the condensation reaction between two amino acids, or an amino acid and a peptide, or a peptide and a peptide can be carried out according to the usual condensation methods such as the axide method, mixed acid anhydride method, DCC(N,N′-dicyclohexylcarbodiimide) or DIC(N,N′-diisopropylcarbodiimide) methods, active ester method, p-nitrophenyl ester method, BOP (benzotriazole-1-yl-oxy-tris [dimethylamino]phosphonium hexafluorophosphate) method, N-hydroxysuccinic acid imido ester method, etc, and Woodward reagent K method.
  • the axide method mixed acid anhydride method
  • active ester method p-nitrophenyl
  • benzyloxycarbonyl (abbreviated Z), isonicotinyloxycarbonyl (iNoc), o-chlorobenzyloxycarbonyl [Z(2Cl) or 2-Cl—Z], p-nitrobenzyloxycarbonyl [Z(NO 2 )], p-methoxybenzyloxycarbonyl [Z(OMe)], t-butoxy-carbonyl (Boc), t-amyloxycarbonyl (Aoc), isobornyloxycarbonyl, adamantyloxy-carbonyl (Adoc), 2-(4-biphenyl)-2-propyloxycarbonyl (Bpoc), 9-fluorenylmethoxycarbonyl (Fmoc), methylsulfonyethoxycarbonyl (Msc), trifluoroacetyl, phthalyl (Pht), formyl (For),
  • side chain protecting groups include acetyl (Ac), benzoyl (Bz), tert butyl (t-Bu), triphenylmethyl (trityl, Trt), tetrahydropyranyl, benzyl (Bzl), 2,6-dichlorobenzyl, nitro, p-toluenesulfonyl (Tos), xanthyl (Xan), benzyl, methyl, ethyl, and t-butyl ester, and aromatic or aliphatic urethan-type protecting groups, photolabile groups such as nitro veratryl oxycarbonyl (Nvoc), and fluoride labile groups such as trimethylsilylethyloxycarbonyl (TEOC).
  • amino terminal protecting groups include: (1) acyl-type protecting groups, such as formyl, acrylyl (Acr), benzoyl (Bz) and acetyl (Ac); (2) aromatic urethan-type protecting groups, such as benzyloxy-carbonyl (Z) and substituted Z, such as p-chlorobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl; (3) aliphatic urethan protecting groups, such as t-butyloxycarbonyl (BOC), diisopropylmethoxycarbonyl, isopropyloxycar-bonyl, ethoxycarbonyl, allyloxycarbonyl; (4) cycloalkyl urethan-type protecting groups, such as 9-fluorenyl-methyloxycarbonyl (Fmoc),
  • Preferred protecting groups include 9-fluorenylmethyloxycarbonyl (Fmoc), 2-(4-biphenylyl)-propyl(2)oxycarbonyl (Bpoc), 2-phyenlpropyl(2)-oxycarbonyl (Poc), and t-butyloxycarbonyl (Boc).
  • Protective groups for the carboxy functional group are exemplified by benzyl ester (OBzl), cyclohexyl ester (Chx), 4-nitrobenzyl ester (ONb), t-butyl ester (Obut), 4-pyridylmeth-yl ester (OPic), and the like. It is often desirable that specific amino acids such as arginine, cysteine, and serine possessing a functional group other than amino and carboxyl groups are protected by a suitable protective group.
  • the guanidino group of arginine may be protected with nitro, p-toluenesulfonyl, benzyloxycarbonyl, adamantyloxycarbonyl, p-meth-oxybenzesulfonyl, 4-methoxy-2,6-dimethylbenzenesulfonyl (Nds), 1,3,5-trimethylphenysul-fonyl (Mts), and the like.
  • the thiol group of cysteine can be protected with p-methoxybenzyl, trityl, and the like.
  • a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt or solvate thereof, together with one or more pharmaceutically carriers thereof and optionally one or more other therapeutic ingredients.
  • the carrier(s) must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
  • the formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous and intraarticular), rectal and topical (including dermal, buccal, sublingual and intraocular) administration.
  • the most suitable route may depend upon the condition and disorder of the recipient.
  • the formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Methods for making such formulations include the step of bringing into association a compound of formula I or a pharmaceutically acceptable salt or solvate thereof (“active ingredient”) with the carrier which constitutes one or more accessory ingredients.
  • active ingredient a pharmaceutically acceptable salt or solvate thereof
  • the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation.
  • Formulations in accordance with embodiments of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion.
  • the active ingredient may also be presented as a bolus, electuary or paste.
  • a tablet may be made by compression or molding, optionally with one or more accessory ingredients.
  • Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, lubricating, surface active or dispersing agent.
  • Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.
  • the tablets may optionally be coated or scored and may be formulated so as to provide sustained, delayed or controlled release of the active ingredient therein.
  • Formulations for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient.
  • Formulations for parenteral administration also include aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickening agents.
  • the formulations may be presented in unit-dose of multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of a sterile liquid carrier, for example saline, phosphate-buffered saline (PBS) or the like, immediately prior to use.
  • Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.
  • Formulations for rectal administration may be presented as a suppository with the usual carriers such as cocoa butter or polyethylene glycol.
  • Formulations for topical administration in the mouth include lozenges comprising the active ingredient in a flavoured basis such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a basis such as gelatin and glycerin or sucrose and acacia.
  • Preferred unit dosage formulations are those containing an effective dose, as hereinbelow recited, or an appropriate fraction thereof, of the active ingredient.
  • formulations in accordance with embodiments of this invention may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavoring agents.
  • compounds in accordance with embodiments of the invention may be used for the treatment or prevention of certain diseases or conditions.
  • preventing refers to administering a medicament beforehand to forestall or obtund an attack.
  • the person of ordinary skill in the medical art recognizes that the term “prevent” is not an absolute term. In the medical art it is understood to refer to the prophylactic administration of a drug to substantially diminish the likelihood or seriousness of a condition, and this is the sense intended in applicants' claims. The reader's attention is directed to the Physician's Desk Reference, a standard text in the field, in which the term “prevent” occurs hundreds of times.
  • FIG. 1 shows antigen-specific increase of the ear thickness derived from the experiment described in Example 2;
  • FIG. 2 shows total numbers of cells in the parotid lymph nodes observed in the experiment described in Example 2;
  • FIG. 3 shows the content of viable and dead cells in the parotid lymph nodes described in Example 2;
  • FIG. 4 presents number and participation of cell types in the draining lymph nodes registered in the experiment described in Example 2;
  • FIG. 5 presents toxicity of the compound of formula IA against mononuclear cells from human blood in as described in Example 3;
  • FIG. 6 presents effects of the intraperitoneal administration of the compound of formula IA on the humoral immune response of mice to sheep erythrocytes as described in Example 4;
  • FIG. 7 shows effects of the intraperitoneal administration of the compound of formula IA on the cellular immune response to ovalbumin as described in Example 5;
  • FIGS. 8A and 8B show the effects of peptides in accordance with embodiments of the invention on PMBC survival in vitro;
  • FIGS. 9A and 9B show the effects of peptides in accordance with embodiments of the invention on PHA-induced PBMC proliferation in vitro;
  • FIGS. 10A and 10B show the effects of several compounds on changes in ear thickness in response to antigenic challenge, as described in Example 7;
  • FIG. 11 shows the permeability of capillary blood vessels in the Evans blue test, as described in Example 7.
  • FIG. 12 shows the total number of cells in the draining lymph nodes, as described in Example 7.
  • FIG. 13 shows the effects of the compounds on the numbers of circulating leukocytes, as described in Example 7;
  • FIG. 14 which shows the types of leukocytes present in different cases, as described in Example 7.
  • FIG. 15 provides morphometric data on the number and composition of cells in mouse auricles, as described in Example 7.
  • Cyclic tetrapeptides according to embodiments of the invention can be synthesized by the use of known peptide synthesis methods, such as solution-phase or solid-phase methods. In general the synthesis involves two consecutive steps: (1) synthesis of a linear tetrapeptide and (2) cyclization to yield the cyclic tetrapeptide.
  • the linear tetrapeptide may be prepared in protected form and then deprotected prior to cyclization.
  • the resin was swelled in dimethylformamide (DMF) (0.25 mmol, 10 ml/g resin) for 15 min. 2. The Fmoc group was removed with a 20% solution of piperidine in DMF (2 ⁇ 20 min). 3. The resin was washed with DMF (3 ⁇ 2 min) 4. The resin was washed with methanol (MeOH) (3 ⁇ 2 min) 5. The resin was washed with dichloromethane (DCM) (3 ⁇ 2 min) 6.
  • DMF dimethylformamide
  • Amino acid or peptide amino groups on the resin were acylated by shaking with a mixture of Fmoc-protected amino acid (4 eq), HBTU or TBTU (4 eq) and DIPEA (4 eq) dissolved in anhydrous DMF (4 ml/mmol), for 20 hours. 7. The resin was washed with DCM (3 ⁇ 2 min) 8. The resin was washed with MeOH (3 ⁇ 2 min) 9. The resin was washed with DCM (3 ⁇ 2 min) 10. The Kaiser test (for all amino acids except proline) was used to estimate if all amino groups were acylated.
  • the resin was washed according to points 3-5 of the protocol.
  • the Fmoc group was removed from the peptide as described in point 2, and the resin was washed again with DMF, MeOH and DCM as in points 3-5.
  • the resin was dried overnight in a desiccator over KOH pellets under reduced pressure at room temperature.
  • Peptides were cleaved from the dried, Wang-type resin with a mixture of trifluoroacetic acid/water/triisopropylsilane 95:2.5:2.5 (v/v/v; 10 ml/1 g of peptidyl-resin).
  • the solution obtained was partially evaporated at room temperature under reduced pressure and peptide was precipitated with 10 volumes of ether. After being filtered off, the crude peptide was dissolved in 0.05 M aqueous HCl and evaporated to dryness. The residue was dissolved in water and lyophilized.
  • Peptides were cleaved from 2-Cl-trityl resin by treatment with a 1:3:1 mixture of acetic acid/dichloromethane/trifluorethanol (v/v/v; 10 ml/1 g of peptidyl-resin).
  • the resulting cleavage solution plus collected washings were filtered, evaporated to dryness at ambient temperature and reduced pressure, and the residue dissolved in a minimal volume of DCM, diluted with 20 volumes of hexane and re-evaporated (twice). Crude detached peptide was dissolved in water and lyophilized.
  • the aim of this experiment was to verify the therapeutic action of compound I-A and its toxicity in a generally accepted animal model.
  • the compound of formula I-A was applied as a therapeutic preparation in the form of 0.1% wt/wt ointment based on a commonly used pharmaceutical vehicle, namely an ointment composed of 50% vaseline and 50% lanoline.
  • a commonly used pharmaceutical vehicle namely an ointment composed of 50% vaseline and 50% lanoline.
  • mice BALB/c female mice, 8-10 week-old, delivered by the Institute of Laboratory Medicine, ód ⁇ , Tru, were used for the study. The mice were fed a commercial, pelleted food and water ad libitum. The local ethics committee approved the study.
  • Cyclic tetrapeptide (compound I-A) was synthesized as described above.
  • Protopic® (tacrolimus) was purchased from Astellas, Ireland; Elidel® (pimecrolimus) was purchased from Novartis; Hydrocortisonum® (hydrocortisone) was purchased from Aflofarm Farmacja Polska, Poland.
  • DMSO was obtained from Fluka; oxazolone, acetone, Evans blue, Giemsa, May-Grünwald, and formalin were from Sigma.
  • mice were shaved on the abdomen (2 ⁇ 2 cm area) and after 24 h 100 ⁇ l of 0.5% oxazolone in acetone was applied to the shaved area.
  • the contact sensitivity reaction was elicited 5 days later by application of 50 ⁇ l of 1% oxazolone in acetone on both sides of the ears. Ear edema was measured 48 h later using a spring caliper.
  • the results were presented as antigen-specific increase of ear thickness (i.e. the background (BG) ear thickness of mice was subtracted from the measured thickness).
  • BG background
  • the compound of formula I-A was applied topically as a 0.1% ointment on both sides of the ears (total volume of 50 ⁇ l per ear), at 24 h and 26 h after elicitation of the reaction with the second dose of oxazolone.
  • Reference compounds were used in a similar fashion in the form of commercially available preparations.
  • mice were subjected to halothane anesthesia and bled from the retro-orbital plexus, followed by cervical dislocation.
  • the number of blood leukocytes was determined by dilution of blood in Matk's solution and counting the cells in a hemocytometer.
  • Blood smears were prepared on microscope glass, dried and stained with Giemsa and May-Grünwald reagents. The smears were subsequently reviewed histologically. The circulating leukocyte numbers were presented per 1 mm 3 and the blood cell compositions as a percentage of a given cell type. Mice treated only with the eliciting dose of antigen served as a background control.
  • the auricles were fixed in 4% formalin solution for 48 h, washed for 24 h, dehydrated in an alcohol series and embedded in paraffin.
  • the paraffin blocks were sliced in a Micron HM310 microtome into 6 ⁇ m sections. The sections were stained with haematoxylin and eosin and with toluidine blue.
  • the histological analysis was performed in a Nikon Eclipse 801 light microscope. On the histological slides containing cross-sections of auricles, the morphometric estimations of neutrophils, macrophages, lymphocytes and mast cells in the perivascular and subepithelial connective tissue were performed. The cells were counted on the area of 0.07 mm 2 at 400 ⁇ magnification. Morphometric analysis was done with the aid of imagine software NIS-Elements (Nikon). For every examined group, 25 enumerations of neutrophils, macrophages, lymphocytes and mast cells were carried out.
  • FIG. 1 presents only antigen-specific increases of the ear thickness (as a result of subtracting background values measured in non-sensitized mice which were given only the eliciting dose of antigen).
  • Compound I-A caused about 80% inhibition of the ear edema; Protopic® and Elidel® respectively caused about 30% and 50% inhibition.
  • FIG. 2 shows that both the compound of formula I-A as well as Elidel® decreased the numbers of lymphocytes in the draining lymph nodes to the level registered in non-sensitized mice. However, in mice treated with Protopic® the number of lymph node cells was similar to that in untreated mice.
  • FIG. 3 shows the proportions of viable and dead cells in the draining lymph nodes, expressed in percentages.
  • the compound of formula I-A exhibits a negligible toxic effect as compared to the control, non-treated mice. A higher toxic effect is caused by Elidel®, and Protopic® is exceptionally toxic with regard to lymph node cells.
  • FIG. 4 depicts the numbers and participation of basic cell types involved in the local inflammatory process.
  • Mast mastocytes
  • L lymphocytes
  • M ⁇ macrophages
  • Ne neutrophils.
  • the auricles from untreated mice (K+) are characterized by a high infiltration of neutrophils.
  • Application of the compound of formula I-A almost entirely reversed the changes observed in control mice (normalization of neutrophil number with some increase in the macrophage content).
  • Protopic® in turn, caused some changes in the proportion of respective cell types with no reduction of the total cell infiltrate. Elidel® caused a moderate diminution of the total cell number.
  • PBMC peripheral mononuclear blood cells
  • CLA cyclolinopeptide
  • Venous blood from a single donor was taken into heparinized syringes, diluted twice with phosphate buffered saline (PBS) and applied onto Lymphoprep® (Polfa, Kutno, Tru) (density of 1.077 g/ml). After centrifugation at 1200 ⁇ g for 20 min, the mononuclear cells from the interphase were harvested and washed 3 times with PBS. The cells were re-suspended in a standard culture medium consisting of RPMI-1640 medium, L-glutamine, sodium pyruvate, 2-mercaptoethanol, 100 ⁇ g/ml each of streptomycin and penicillin, and 10% fetal calf serum.
  • PBS phosphate buffered saline
  • the cells were distributed to in 96-well flat-bottom culture plates at density of 2 ⁇ 10 5 /100 ⁇ l.
  • the compounds (formula I-A and CLA) were initially dissolved in DMSO (5 mg/500 ⁇ l) and subsequently in the culture medium.
  • DMSO appropriately diluted with the culture medium, was used as a control.
  • cell viability was determined by a colorimetric method (Hansen et al., J Immunol Methods, 1989, 119, 203-210).
  • FIG. 5 presented as mean optical density values from quadruplicate wells (cell cultures) ⁇ SE.
  • the compound of formula I-A (listed as “4B8M”) did not show appreciable toxicity in the concentration range of 10 to 100 ⁇ g/ml.
  • CLA on the other hand, showed a statistically significant cytotoxic effect at 40 ⁇ g/ml.
  • mice CBA female mice, 8-12 weeks old, were delivered by The Institute of Laboratory Medicine, ód ⁇ , Tru. The mice had free access to water and pelleted food. The local ethics committee approved the study. Sheep erythrocytes (SRBC) were delivered by Wroclaw University of Life and Environmental Sciences, Poland, and maintained on RPMI-1640 medium.
  • SRBC Sheep erythrocytes
  • mice were immunized with 0.2 ml of 5% SRBC suspension (0.5 ml of SRBC pellet re-suspended to a volume of 10 ml of 0.9% NaCl), intraperitonelly. After 4 days the number of antibody-forming cells (AFC) in the spleens was determined using an assay of local hemolysis in agar gel (per Mishell et al., J Exp Med, 1967, 126, 423-442). The results are presented in FIG. 6 as a mean value of 5 mice ⁇ standard error and expressed as AFC number per 10 6 of viable splenocytes.
  • AFC antibody-forming cells
  • mice were immunized with SRBC as described above and after 2 h were given 10 or 100 ⁇ g of the compound of formula I-A.
  • Cyclosporin A (CsA) served as a reference drug.
  • the number of antibody-forming cells to SRBC was measured after 4 days.
  • the compound of formula IA was more inhibitory at both doses than CsA.
  • mice 8-12 weeks old were delivered by The Institute of Laboratory Medicine, ód ⁇ , Tru.
  • the mice had free access to water and pelleted food.
  • Ovoalbumin was from Sigma and the adjuvants from Difco.
  • mice were sensitized subcutaneously with 5 ⁇ g of ovalbumin (OVA) emulsified in Freund's complete adjuvant in the tail base. After 4 days the mice were challenged with 50 ⁇ g of OVA in Freund's incomplete adjuvant in the hind footpads. Following the next 24 hours, the footpad thickness was measured using a caliper. Controls (background response mice) were not sensitized but received the challenging dose of OVA.
  • the compound of formula I-A and the reference compound were administered to mice in two 100 ⁇ g intraperitoneal doses, 2 h before and 24 h after the sensitizing dose of antigen. The results, presented in FIG.
  • Cyclic tetrapeptides were tested in vitro for their effects on phytohemagglutinin A (PHA)-induced proliferation of human peripheral blood mononuclear cells (PBMC) and for lipopolysaccharide (LPS)-induced production of tumor necrosis factor alpha (TNF- ⁇ ) production by whole blood cell cultures at 1-100 ⁇ g/ml concentration range. Compounds were also tested for cell toxicity at 1-100 ⁇ g/ml concentration range against human PBMC.
  • PHA phytohemagglutinin A
  • PBMC peripheral blood mononuclear cells
  • LPS lipopolysaccharide
  • TNF- ⁇ tumor necrosis factor alpha
  • RPMI-1640 medium Cibi/Life Technologies, UK
  • FCS fetal calf serum
  • FCS Gibco
  • DMSO phytohemagglutinin A
  • LPS lipopolysaccharide
  • MTT 93-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
  • SDS SDS and DMF
  • the culture medium consisted of RPMI-1640, 10% addition of FCS, L-glutamine, sodium pyruvate, 2-mercaptoetanol and antibiotics (streptomycin and penicillin). Cyclic tetrapeptides were initially dissolved in DMSO (5 mg/ml), the dissolved in the culture medium to the desired concentration.
  • Venous blood was taken from a single donor (a male, 62-years old) into heparinized syringes and diluted twice with phosphate-buffered saline (PBS).
  • PBMC phosphate-buffered saline
  • PBMC peripheral blood mononuclear cells
  • the interphase cells consisting of lymphocytes (20%) and monocytes (80%) were then washed three times with Hanks' medium and re-suspended in the culture medium at density of 2 ⁇ 10 6 cells/ml.
  • the isolated PBMC were distributed into 96-well flat-bottom plates in 100 ⁇ l aliquots (2 ⁇ 10 5 cells/well).
  • PHA was used at concentration of 5 ⁇ g/ml.
  • the compounds were tested at concentrations of 1, 10 and 100 ⁇ g/ml.
  • DMSO at appropriate dilutions served as control.
  • the proliferative response of cells was determined by the colorimetric MTT method (Hansen et al., J. Immunol. Methods, 1989, pp. 203-210).
  • the data are presented as a mean OD value from quadriplicate wells ⁇ standard error (SE).
  • the cultures “Control ( ⁇ )” contained no mitogen (PHA).
  • the cultures “Control (PHA)” contained PHA but not cyclic tetrapeptides.
  • PBMC peripheral blood mononuclear cells
  • PBMC peripheral blood mononuclear cells
  • the assay was performed per Hansen et al., J. Immunol. Methods, 1989, 119 pp. 203-210. Briefly, 25 ⁇ l of MTT (5 mg/ml) stock solution was added per well at the end of cell incubation and the plates were incubated for 3 h in a cell culture incubator. Then, 100 ⁇ l of the extraction buffer (20% SDS with 50% DMF, pH 4.7) was added. After additional overnight incubation, the optical density was measured at 550 nm (Dynatech 5000).
  • results are presented as mean values ⁇ standard error (SE).
  • Brown-Forsyth's test was used to determine the homogeneity of variance between groups. When the variance was homogenous, analysis of variance (one-way ANOVA) was applied, followed by post hoc comparisons with the Tukey's test to estimate the significance of the differences between groups. Significance was determined at P ⁇ 0.05. Statistical analysis was performed using STATISTICA 6.1 for Windows.
  • FIGS. 8A and 8B The effects of the peptides on PMBC survival in 24 h culture are presented in FIGS. 8A and 8B .
  • Peptide 4B8M (compound I-A) was included as a reference compound. Appropriate dilutions of DMSO were added to control cultures. The results showed no signs of toxicity of the compounds in the studied concentration range.
  • P01 compound I-D
  • P02 compound I-E
  • P03 compound I-F
  • P04 compound I-G
  • P05 compound I-H
  • P06 compound I-J
  • P07 compound I-K
  • P08 compound I-L
  • P10a compound I-M
  • P10b compound I-N
  • P11 compound I-O.
  • FIGS. 8A and 8B show the effects of the tested peptides on the survival of PBMC.
  • FIGS. 9A and 9B The effects of the peptides on the proliferative response of PMBC are presented in FIGS. 9A and 9B .
  • Peptide 4B8M was included as a reference compound. Appropriate dilutions of DMSO were added to control cultures.
  • mice 8-10 weeks old, were delivered by the Institute of Laboratory Medicine, ód ⁇ , Tru. The mice were fed a commercial, pelleted food and water ad libitum. The local ethics committee approved the study.
  • Compound I-A was synthesized as described above; Protopic® (tacrolimus) was from Astellas, Ireland; Elidel® (pimecrolimus) from Novartis; DMSO from Fluka; TDI, acetone, Evans blue, Trypan blue, Giemsa, May-Grünwald, haematoxylin, eosin, toluidine blue and formalin were from Sigma.
  • the test was performed according to Yamamoto, Eur. J. Pharmacol., 2006, 550, 166-172, with minor modifications. Mice were shaved on the abdomen (2 ⁇ 2 cm area) and after 24 h 100 ⁇ l of 3% TDI in acetone was applied through 3 consecutive days. After 14 days the reaction was elicited by application of 50 ⁇ l of 0.3% TDI on both sides of the ears. The procedure was repeated 5 times every 3 days. Ear thickness was measured using a spring caliper (Mitutoyo) 5 h and 24 h after each challenge with TDI.
  • Compound I-A was applied topically in the form of 0.1% ointment on both sides of the ears (total volume of 100 ⁇ l ⁇ 50 ⁇ l per ear), one hour after each challenge with TDI.
  • the reference drugs were applied in a similar way.
  • mice were subjected to halothane anesthesia and bled from the retro-orbital plexus, followed by the cervical dislocation.
  • the number of blood leukocytes was determined by dilution of blood in Matk's solution and counting the cells in a hemocytometer.
  • Blood smears were prepared on microscope glass, dried and stained with Giemsa and May-Grünwald reagents. The smears were subsequently reviewed histologically. The cell numbers were presented per 1 ⁇ l and the blood cell compositions as a percentage of a given cell type.
  • mice were given 1 mg of Evans blue in 0.2 ml of 0.9% NaCl, intravenously. After 30 min mice were sacrificed, the ears were cut off, weighed and immersed in 50 ⁇ l of 1M KOH for 18 h at 37° C. The dye was extracted from the ears using 450 ⁇ l of 0.2 M phosphate acid and acetone (5:13 ratio). The samples were centrifuged at 3,000 rpm for 15 min. The optical densities (OD) of the supernatants were measured at 630 nm. The amount of Evans blue ( ⁇ g/ml) was determined based on a standard curve. The results were presented as the amount of Evans blue per 100 mg of wet tissue. Mice treated only with the eliciting dose of antigen served as a background control.
  • the auricles were fixed in 4% formalin solution for 48 h, washed for 24 h, dehydrated in an alcohol series and embedded in paraffin.
  • the paraffin blocks were sliced in a Micron HM310 microtome into 6 ⁇ m sections. The sections were stained with haematoxylin and eosin and with toluidine blue. Histological analysis was performed using a Nikon Eclipse 801light microscope. Morphometric estimations of neutrophils, macrophages, lymphocytes and mast cells in the perivascular and subepithelial connective tissue were performed on the histological slides containing cross-sections of auricles. Cells were counted on an area of 0.07 mm 2 at 400 ⁇ magnification. Morphometric analysis was performed using an imagine software NIS-Elements (Nikon). For every preparation examined, 25 enumerations of neutrophils, macrophages, lymphocytes and mast cells were carried out.
  • FIGS. 10A and 10B are presented as mean values ⁇ standard error (SE). Brown-Forsyth's test was used to determine the homogeneity of variance between the groups. When the variance was homogenous, analysis of variance (one-way ANOVA) was applied, followed by post hoc comparisons with the Tukey's test to estimate the significance of the differences between groups. Nonparametric data were evaluated with the Kruskal-Wallis' analysis of variance, as indicated in the text. Significance was determined at P ⁇ 0.05. Statistical analysis was performed using STATISTICA 7 for Windows.
  • FIGS. 10A and 10B show ear thickness measure 5 h ( FIG. 10A ) and 24 h ( FIG. 10B ) after administration on the day of the test indicated in the figure.
  • Control responses to TDI gradually elevated after each antigen challenge (best seen in the 5 h measurement). The results showed differentiated efficacy of the compounds in reducing the ear swelling.
  • FIG. 11 shows the permeability of capillary blood vessels in the Evans blue test.
  • the rates of blood vessel permeability were strictly correlated with the effects of the compounds on ear thickness in the respective mouse groups.
  • FIG. 12 shows the total number of cells in the draining lymph nodes.
  • the treatment of mice with compound I-A resulted in a reduction of the lymph node cell numbers almost to the background levels (non-sensitized mice).
  • FIG. 13 shows the effects of the compounds on the numbers of circulating leukocytes; the application of the studied preparations lowered the numbers of circulating leukocytes to the levels observed in control, unsensitized mice.
  • the blood composition in control mice with fully developed reaction to TDI was characterized by an increased content of neutrophils and eosinophils compared to control, background mice ( FIG. 14 , which shows a breakdown of the types of leukocytes in each case).
  • the blood picture was normalized upon application of 4B8M (I-A) (a reduction of neutrophil and eosinophil contents) but not following administration of Protopic® or Elidel®.
  • FIG. 15 provides morphometric data on the number and composition of cells in the auricles.
  • the composition of cell types within the auricles differed among the studied mouse groups is presented in FIG. 15 .
  • the predominant, residing cell types in control non-sensitized mice are mastocytes and neutrofils (10 and 5 cells per the analyzed area, respectively).
  • the number of mastocytes increased twofold and neutrophils almost 5-fold (20 and 23 cells, respectively).
  • Protopic® and Compound I-A were effective in reducing the cell numbers to 14 and 14.8.
  • the cells were then washed twice with Hanks' medium, passed through glass wool column to remove debris, and re-suspended in the culture medium, referred to below as the “culture medium”, consisting of RPMI-1640, supplemented with 10% of fetal calf serum, L-glutamine, sodium pyruvate, 2-mercaptoethanol, streptomycin and penicillin (100 ⁇ g/ml).
  • the cells were then distributed into 96-well flat-bottom tissue culture plates (Nunc) at a density of 2 ⁇ 10 5 cells/100 ⁇ l/well.
  • SRBC Sheep Erythrocytes
  • mice were sensitized intraperitoneally with 0.2 ml of 5% (v/v) SRBC suspension. After four days spleens from these mice were isolated and single cell suspensions prepared by homogenization in PBS solution. After washing the cells in PBS by centrifugation, the cells were re-suspended in the culture medium at a density of 5 ⁇ 10 6 cells/ml. The cells were subsequently distributed to 24-well culture plates in 1 ml aliquots and 0.05 ml of 0.005% SRBC was added as the antigen to each well. The compounds were added to the cultures at the beginning of the four-day incubation period at concentration ranges of 1-100 ⁇ g/ml. The number of antibody-forming cells (AFC) in the cultures was determined using a method of local hemolysis in agar gel according to Mishell et al., J Exp Med, 1967, 126, 423-442.
  • AFC antibody-forming cells
  • Splenocytes at a density of 2 ⁇ 10 5 cells/100 ⁇ l/well, re-suspended in the culture medium, were cultured for 24 h in a cell culture incubator with the compounds (1-100 ⁇ g/ml).
  • Cell survival was determined by MTT colorimetric method. The results were presented as mean optical density (OD) at 550 nm from 4 wells. The viability of cells in respective compound concentrations was compared to appropriate DMSO control groups (100% survival), corresponding to respective compound concentrations.
  • compound I-C showed a strong inhibitory effect on concanavalin A-induced mouse splenocyte proliferation. At 100 ⁇ g/m, this compound showed 70% toxicity to splenocytes. At 10 ⁇ g/m and 100 ⁇ g/m, compound I-C showed 33% and 80% suppression, respectively, in the model of in vitro humoral immune response to SRBC in mouse splenocyte cultures. In the model of delayed type hypersensitivity to ovalbumin, compound I-C showed 26.9% inhibition at the dose of 100 ⁇ g, compared to 72.7% inhibition by compound I-A.
  • compound I-B demonstrated a weak antiproliferative effect on concanavalin A-induced splenocyte proliferation; no such effect was observed at lower concentrations. Compound I-B had 30% toxicity at this concentration.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Immunology (AREA)
  • Epidemiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Dermatology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Pain & Pain Management (AREA)
  • Transplantation (AREA)
  • Rheumatology (AREA)
  • Pulmonology (AREA)
  • Psychology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
US13/579,933 2010-02-19 2011-02-19 Cyclic tetrapeptides and therapeutic applications thereof Abandoned US20130224232A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US13/593,716 US9382292B2 (en) 2010-02-19 2012-08-24 Cyclic tetrapeptides and therapeutic applications thereof

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
PL390493A PL219569B1 (pl) 2010-02-19 2010-02-19 Cykliczne tetrapeptydy i ich zastosowanie
PLP.390493 2010-02-19
PCT/US2011/025571 WO2011103524A2 (en) 2010-02-19 2011-02-19 Cyclic tetrapeptides and therapeutic applications thereof

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2011/025571 A-371-Of-International WO2011103524A2 (en) 2010-02-19 2011-02-19 Cyclic tetrapeptides and therapeutic applications thereof

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US13/593,716 Continuation-In-Part US9382292B2 (en) 2010-02-19 2012-08-24 Cyclic tetrapeptides and therapeutic applications thereof

Publications (1)

Publication Number Publication Date
US20130224232A1 true US20130224232A1 (en) 2013-08-29

Family

ID=44483608

Family Applications (3)

Application Number Title Priority Date Filing Date
US13/579,933 Abandoned US20130224232A1 (en) 2010-02-19 2011-02-19 Cyclic tetrapeptides and therapeutic applications thereof
US13/593,716 Expired - Fee Related US9382292B2 (en) 2010-02-19 2012-08-24 Cyclic tetrapeptides and therapeutic applications thereof
US15/201,565 Active US10238713B2 (en) 2010-02-19 2016-07-04 Cyclic tetrapeptides and therapeutic applications thereof

Family Applications After (2)

Application Number Title Priority Date Filing Date
US13/593,716 Expired - Fee Related US9382292B2 (en) 2010-02-19 2012-08-24 Cyclic tetrapeptides and therapeutic applications thereof
US15/201,565 Active US10238713B2 (en) 2010-02-19 2016-07-04 Cyclic tetrapeptides and therapeutic applications thereof

Country Status (13)

Country Link
US (3) US20130224232A1 (enExample)
EP (1) EP2536732B1 (enExample)
JP (1) JP5901543B2 (enExample)
KR (1) KR101704665B1 (enExample)
CN (1) CN102858779B (enExample)
AU (1) AU2011217882B2 (enExample)
BR (1) BR112012020833A2 (enExample)
CA (1) CA2827694C (enExample)
ES (1) ES2611663T3 (enExample)
IL (2) IL221535A (enExample)
PL (1) PL219569B1 (enExample)
RU (1) RU2568436C2 (enExample)
WO (1) WO2011103524A2 (enExample)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10154997B2 (en) 2015-08-04 2018-12-18 Washington University Treatment of parasitic diseases using KDAC inhibitor compounds
US10238713B2 (en) 2010-02-19 2019-03-26 Lodz University Of Technology Cyclic tetrapeptides and therapeutic applications thereof
US10421778B2 (en) 2015-05-05 2019-09-24 Washington University Isoform-selective lysine deacetylase inhibitors

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018232285A1 (en) * 2017-06-16 2018-12-20 University Of Florida Research Foundation, Incorporated Macrocyclic peptides and derivatives thereof with opioid activity

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010006267A2 (en) * 2008-07-11 2010-01-14 University Of Kansas Cyclic tetrapeptides

Family Cites Families (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3862925A (en) * 1973-07-05 1975-01-28 American Home Prod Preparation of somatotropin release inhibiting factor and intermediates therefor
US3842067A (en) 1973-07-27 1974-10-15 American Home Prod Synthesis of(des-asn5)-srif and intermediates
JPS5726506B2 (enExample) * 1974-03-08 1982-06-04
IT1077010B (it) * 1976-09-01 1985-04-27 Snam Progetti Processo per la preparazione di copolimeri dell isobutilene contenenti insaturazioni ed atomi di alogeno reattivi
US4105603A (en) 1977-03-28 1978-08-08 The Salk Institute For Biological Studies Peptides which effect release of hormones
JPH0334991A (ja) * 1989-06-30 1991-02-14 Shionogi & Co Ltd 環状テトラペプチドおよびその製造法
US5216124A (en) 1989-12-15 1993-06-01 G. D. Searle & Co. Substituted cyclic tetrapeptides
US5541287A (en) 1992-06-09 1996-07-30 Neorx Corporation Pretargeting methods and compounds
US6777217B1 (en) * 1996-03-26 2004-08-17 President And Fellows Of Harvard College Histone deacetylases, and uses related thereto
US6013763A (en) 1996-06-04 2000-01-11 Genentech, Inc. Peptide variants of protein A
NZ507083A (en) * 1998-03-23 2003-10-31 Trimeris Inc Methods and compositions for peptide synthesis, in particular T-20 (DP-178) and T-20-like peptides
US6281331B1 (en) 1998-03-23 2001-08-28 Trimeris, Inc. Methods and compositions for peptide synthesis
DE19816922A1 (de) 1998-04-16 1999-10-21 Wolfgang Beck Einfache Synthese von cyclischen Tetrapeptiden aus nichtaktivierten Peptidestern an Metallzentren
AUPP505798A0 (en) * 1998-08-04 1998-08-27 Fujisawa Pharmaceutical Co., Ltd. Novel compound fr225497 substance
IL142531A0 (en) 1998-10-13 2002-03-10 Fujisawa Pharmaceutical Co Cyclic peptide compound, processes for the preparation thereof and pharmaceutical compositions containing the same
GB9903087D0 (en) * 1999-02-12 1999-03-31 Glaxo Group Ltd Process
ATE376556T1 (de) * 2003-12-31 2007-11-15 Hoffmann La Roche Verfahren zur peptidsynthese unter verwendung einer reduzierten menge an entschützungsmittel
WO2006049597A1 (en) 2004-10-27 2006-05-11 Avanir Pharmaceuticals Amino acid-derived compounds as modulators of the reverse cholesterol transport
US20080219962A1 (en) * 2004-12-14 2008-09-11 Biorunx Co., Ltd. Method to Enhance the Bone Formation Activity of Bmp by Runx2 Acetylation
RU2450018C2 (ru) 2006-08-23 2012-05-10 Йеда Рисерч Энд Диверлопмент Ко. Лтд Конъюгаты rgd-пептидов и фотосенсибилизаторов порфирина или (бактерио)хлорофилла и их применение
DK2257309T3 (da) 2008-02-27 2020-01-20 Yeda Res And Development Company Ltd Rgd-(bakterie)chlorophyl-konjugater til anvendelse ved diagnose af tumorer, der omfatter nekrotiske domæner
GB0918579D0 (en) 2009-10-22 2009-12-09 Imp Innovations Ltd Gadd45beta targeting agents
PL219569B1 (pl) 2010-02-19 2015-05-29 Peptaderm Spółka Z Ograniczoną Odpowiedzialnością Cykliczne tetrapeptydy i ich zastosowanie

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010006267A2 (en) * 2008-07-11 2010-01-14 University Of Kansas Cyclic tetrapeptides

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
Datta et al., Cancer Res. (Dec 1, 2009), Vol:69(23):8902-9, (online Nov. 10, 2009) *
Davies, The Cyclization of Peptides and Depsipeptides, Journal of Peptide Science, vol 9:471-501 (2003). *
Liu et al., Bioorg Med Chem. (Feb 1, 2009), vol:17(3): 1026-1033. *
Morton et al., Calcineurin inhibitors cause an acceleration of the neurological phenotype in a mouse transgenic for the human Huntington's disease mutation, Brain Res Bull. (Apr 18, 2006) Vol:69(6):669-79 *
Norgren et al., beta2-amino acids in the Design of Conformationally Homogeneous cyclo-Peptide Scaffolds, J. Org. Chem., (2006), Vol:71:6814-6821 *
Siemion et al, Cyclolinopeptides and Their Analogs -a New Family of Peptide Immunosuppressants Affecting the Calcineuriin System, Archivum Immunologiae et Therapiae Experimentalis, 1999, Vol 47:143-153 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10238713B2 (en) 2010-02-19 2019-03-26 Lodz University Of Technology Cyclic tetrapeptides and therapeutic applications thereof
US10421778B2 (en) 2015-05-05 2019-09-24 Washington University Isoform-selective lysine deacetylase inhibitors
US10154997B2 (en) 2015-08-04 2018-12-18 Washington University Treatment of parasitic diseases using KDAC inhibitor compounds

Also Published As

Publication number Publication date
PL390493A1 (pl) 2011-08-29
BR112012020833A2 (pt) 2017-12-19
AU2011217882B2 (en) 2015-02-05
US20130108655A1 (en) 2013-05-02
US9382292B2 (en) 2016-07-05
WO2011103524A3 (en) 2011-12-29
KR101704665B1 (ko) 2017-02-08
KR20130088004A (ko) 2013-08-07
RU2568436C2 (ru) 2015-11-20
JP2013520440A (ja) 2013-06-06
US10238713B2 (en) 2019-03-26
JP5901543B2 (ja) 2016-04-13
WO2011103524A2 (en) 2011-08-25
EP2536732A4 (en) 2014-04-30
AU2011217882A1 (en) 2012-10-11
CA2827694A1 (en) 2011-08-25
US20170157204A1 (en) 2017-06-08
WO2011103524A8 (en) 2012-09-13
EP2536732B1 (en) 2016-10-19
ES2611663T3 (es) 2017-05-09
IL221535A (en) 2016-08-31
CN102858779A (zh) 2013-01-02
RU2012139668A (ru) 2014-03-27
CA2827694C (en) 2018-09-18
PL219569B1 (pl) 2015-05-29
IL224410A (en) 2017-07-31
CN102858779B (zh) 2016-08-03
EP2536732A2 (en) 2012-12-26

Similar Documents

Publication Publication Date Title
AU2010310665B2 (en) Peptidic GLP-2 agonists
US10278957B2 (en) Opioid agonist peptides and uses thereof
US9260480B2 (en) Process for the manufacture of Degarelix and its intermediates
US10238713B2 (en) Cyclic tetrapeptides and therapeutic applications thereof
CZ372698A3 (cs) Substituované imidazolidinové deriváty, způsob jejich přípravy, jejich použití a farmaceutické prostředky, které je obsahují
SK166798A3 (en) Peptide derivatives, process for their preparation, pharmaceutical composition them containing, and their use
CZ111997A3 (en) Peptides exhibiting immunomodulating activity, process of their preparation, pharmaceutical compositions containing thereof and their use
SK114998A3 (en) Peptide derivatives, preparation method and use thereof
WO2022177878A1 (en) Glucagon-like peptide-1 receptor antagonists
JP3568952B2 (ja) ５位及び６位に修飾アミノアシル残基を有するｌｈｒｈ拮抗薬
CN105636975B (zh) 抗炎三肽
US9115179B2 (en) Synthesis of beta-turn peptidomimetic cyclic compounds
KR20060073618A (ko) 신규 항종양 화합물
EP0101929A1 (en) Polypeptide-diesters, their production and use
AU671118B2 (en) Tachykinin antagonist tricyclic compounds, preparation of same and pharmaceutical compositions containing such compounds
US20060189539A1 (en) Novel GnRH analogues with antitumour effects and pharmaceutical compositions thereof
HUT63856A (en) Process for producing peptides for treating the ageing and disorders of the immune system and central nervous system
CN117645651A (zh) 一种环肽类抗肿瘤活性化合物及药物组合物与应用
Albericio et al. Kahalalide F and related compounds

Legal Events

Date Code Title Description
STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION

AS Assignment

Owner name: LODZ UNIVERSITY OF TECHNOLOGY, POLAND

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:ZABROCKI, JANUSZ;ZIMECKI, MICHAL;KASZUBA, ANDRZEJ;AND OTHERS;SIGNING DATES FROM 20170731 TO 20171102;REEL/FRAME:044178/0166

Owner name: PEPTADERM SP. Z O.O., POLAND

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:ZABROCKI, JANUSZ;ZIMECKI, MICHAL;KASZUBA, ANDRZEJ;AND OTHERS;SIGNING DATES FROM 20170731 TO 20171102;REEL/FRAME:044178/0166