WO2003017920A2 - Facteurs de protection contre des inflammations, des brulures et des stimuli nocifs - Google Patents

Facteurs de protection contre des inflammations, des brulures et des stimuli nocifs Download PDF

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Publication number
WO2003017920A2
WO2003017920A2 PCT/IL2002/000713 IL0200713W WO03017920A2 WO 2003017920 A2 WO2003017920 A2 WO 2003017920A2 IL 0200713 W IL0200713 W IL 0200713W WO 03017920 A2 WO03017920 A2 WO 03017920A2
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Prior art keywords
gly
stimuli
ala
disease
peptide
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PCT/IL2002/000713
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English (en)
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WO2003017920A3 (fr
Inventor
Uri Wormser
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Yissum Research Development Company Of The Hebrew University Of Jerusalem
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Application filed by Yissum Research Development Company Of The Hebrew University Of Jerusalem filed Critical Yissum Research Development Company Of The Hebrew University Of Jerusalem
Priority to AU2002329017A priority Critical patent/AU2002329017A1/en
Priority to EP02765303A priority patent/EP1572069A4/fr
Publication of WO2003017920A2 publication Critical patent/WO2003017920A2/fr
Priority to US10/790,888 priority patent/US7238656B2/en
Publication of WO2003017920A3 publication Critical patent/WO2003017920A3/fr
Priority to US11/750,408 priority patent/US7605132B2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/36—Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells
    • 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
    • A61K38/10—Peptides having 12 to 20 amino acids

Definitions

  • the present invention relates to pharmaceutical compositions comprising peptides and their derivatives, and to the use of these compositions for treatment of tissue lesions or diseases caused by, or associated with inflammatory processes including, but not limited to, those resulting from chemical bums, thermal burns and additional noxious stimuli.
  • peptides useful in the treatment of malignancies and bacteriological infections have been disclosed, nowhere in the background art is it taught or suggested that peptides produced and/or released by skin in response to thermal injury after treatment with iodine preparations may be useful for the prevention of tissue trauma or damage induced by chemical or thermal insults.
  • peptide factors isolated from skin following exposure to chemical or thermal injury and treatment with iodine preparations are capable of reducing or ameliorating the extent of injury if administered to the skin of other animals. It is emphasized that while the principles of the invention are exemplified herein below through the use of animal models the invention is useful for humans, and that the human homologs of all disclosed peptides are explicitly included within the scope of the invention.
  • the peptides isolated from the skin are selected from the group consisting of: SEQ ID NO: l
  • Peptide HI was found to be a partial sequence of Histone H2A with its carboxy terminal Gly was substituted by Ala.
  • Peptide VI was identified as guinea pig fibrinopeptide A, while peptides TV and VII differ from peptide VI by omission and addition, respectively, of an amino terminal threonine.
  • the peptides may be elongated or truncated by one or more residues without altering their basic protective attributes. Active fragments, as well as deletions or substitutions as well as derivatives are also possible without detracting from the activity of the specific disclosed peptides.
  • any known peptides such as Fibrinopeptide A designated herein by SEQ ID NO: 3 or SEQ ID NO: 9, are excluded from the novel compounds, but are disclosed and claimed for all of the novel uses disclosed herein. Methods of using these peptides are also disclosed, comprising administering to an individual in need thereof a therapeutically effective amount of a pharmaceutical composition comprising as an active ingredient a peptide according to the invention.
  • a pharmaceutical composition comprising as an active ingredient a peptide according to the invention.
  • the individual is a human subject and the peptide is a human homolog or analog of the sequences disclosed herein.
  • the pharmaceutical compositions may further comprise at least one additional agent or drug, including but not limited to known anti-inflammatory agents, or chemokine/cytokine modulatory agents.
  • compositions according to the present invention will be useful for treating a diverse group of indications having an inflammatory or autoimmune mechanism involved in their etiology or pathogenesis exemplified by psoriasis, systemic lupus erythematosus (SLE), multiple sclerosis, inflammatory bowel disease (Crohn's disease) and arthritis, including rheumatoid arthritis, asthma, and certain degenerative diseases including amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease and muscular dystrophy, as well as sepsis and malignant or benign tumors having inflammatory component or involving inflammatory processes.
  • compositions of the invention are particularly useful for preventing, reducing or treating the tissue damage consequent to trauma or other noxious insults including but not limited to heat stimuli, cold stimuli, chemical stimuli, electric stimuli, ultraviolet irradiation, ionizing and non-ionizing irradiation, and ultrasound.
  • Fig. 1 shows the protective effect of the skin extract against thermal skin burns.
  • Fig. 2 shows the protective effect of the mixture of peptides ILT, TV, VI, and VII against thermal or chemical skin burns.
  • Fig. 3 depicts the protective effect of each of the peptides III, TV, VI, or VII against chemical skin burns.
  • Fig. 4 depicts the protective effect of each of the peptides HI, TV, VI, or VII against thermal skin burns.
  • Fig. 5 shows the effect of the mixture of peptides III, IV, VI, and VIT on body weight loss induced by sulfur mustard.
  • Fig. 6 shows the protective effect of N-methylated analogs of peptide TJI against skin burns induced by sulfur mustard.
  • Fig. 7 demonstrates the protective effect of peptides III, 3b and TV against SM in the mouse inflammation (ear edema) model.
  • Fig. 8 shows the protective effect of anti mouse interleukin 1 beta (IL1 beta) antibodies, anti mouse tumor necrosis factor alpha (TNF alpha) antibodies, peptides 3m, 3ml and IV against SM in the mouse ear edema model.
  • IL1 beta interleukin 1 beta
  • TNF alpha tumor necrosis factor alpha
  • the present invention relates to peptides and their derivatives capable of protecting tissue against noxious stimuli. More particularly, the present invention provides peptides and their derivatives that, by administration to the body, reduce the degree, or prevents the development of lesions caused by many kinds of noxious stimuli including heat, one or more chemicals, irradiation and combinations of these stimuli.
  • the inventor has previously demonstrated that topical application of iodine or povidone iodine preparations immediately after exposure to heat reduces, and many times nullifies, the burning sensation and more importantly, prevents or significantly reduces, skin damage that was expected to develop without iodine treatment [Wormser, U. (1998) Bums 24, 383].
  • iodine preparations were also demonstrated against mustard gas (sulfur mustard) and non-mustard vesicants. It was shown that post-exposure treatment with topical iodine preparations significantly reduced the degree of skin lesion caused by the chemical agents [Wormser U et al. (1997) Arch. Toxicol. 71, 165-170; Wormser U et al. (2000) Toxicol. Appl. Pharmacol. 169, 33-39].
  • guinea pig skin was exposed to heat and immediately thereafter applied with iodine. Extracts of the skin were injected intradermally into new, naive guinea pigs. Immediately thereafter the guinea pigs were exposed to heat stimuli; the animals were checked for gross pathology during the following days.
  • Figure 1 demonstrates the protective effect of the extract against thermal bums. In the animal group treated with the extract there was a statistically significant reduction of 68% in the ulceration area as compared to the control animals injected with saline.
  • the present invention now identifies peptide factors that are isolated from the skin after exposure to thermal or chemical injury followed by treatment with iodine. These peptides have been identified, prepared synthetically in substantially pure form, and shown to confer their protective effects on the skin of naive animals.
  • the present invention relates to protection by the factors against inflammation and inflammatory processes including but not limited to skin bums.
  • Bums relate to tissue damage caused by heat stimuli, cold stimuli, chemical stimuli, electric stimuli, ultraviolet irradiation, ionizing and non-ionizing irradiation, irradiation of all kinds including electromagnetic and ultrasound.
  • Histone H2A fragment 36-44 peptide HI and its derivatives including the derivatives of the peptide 3 series
  • Histone H2A fragment 36-44 are species dependent sequences, and related sequences, especially human homologues are explicitly included in the scope of the invention. All active analogs of these sequences are also within the scope of the invention.
  • Active analogs may encompass many variants as arc well known in the art, including but not limited to truncations or extensions of amino acids at the amino terminus or carboxy terminus, insertion or deletion of amino acids, N-methylated analogs, and other modifications, provided that these analogs possess anti- inflammatory properties.
  • the anti-inflammatory properties of Fibrinopeptide A (peptide IV and its derivatives), are also species dependent sequence, and related sequences especially human homologues are explicitly included in the scope of the invention. All active analogs of these sequences are also within the scope of the invention.
  • Active analogs may encompass many variants as are well known in the art, including but not limited to truncations or extensions of amino acids at the amino terminus or carboxy terminus, insertion or deletion of amino acids, N-methylated analogs, and other modifications, provided that these analogs possess anti-inflammatory properties
  • fibrinopeptide A fragments are synergistically enhanced when given together with another anti-inflammatory peptide of the present invention, e.g., peptide 3ml , or with other known anti-inflammatory agents, e.g., anti TNF alpha antibodies and anti IL1 antibodies.
  • another anti-inflammatory peptide of the present invention e.g., peptide 3ml
  • other known anti-inflammatory agents e.g., anti TNF alpha antibodies and anti IL1 antibodies.
  • noxious stimuli as used herein and in the claims is intended to encompass any trauma or insult or disease or condition involving an inflammatory response, since the toxicity of SM results from inflammatory response and involvement of inflammatory mediators, namely, induction, synthesis, activation, secretion are involved in the evolution and creation of tissue damage caused by noxious stimuli (thermal, chemical, irradiation, etc.). Since the peptides have proven to be particularly efficacious against skin bums, they were also tested and demonstrated to act as bona fide anti-inflammatory agents.
  • the peptides are expected to be efficacious in all diseases or disorders that involve inflammation or inflammatory activity. Therefore, this invention relates to the protective effect of the H2A histone 36-44 fragment and its analogs, and fibrinopeptide A against all disorders or diseases that are related to or involve inflammatory process (whether alone or together with additional known anti-inflammatory agents, including but not limited to TNF and IL1 antibodies).
  • inflammatory processes related to diseases, e.g., psoriasis, arthritis, Crohn's disease, degenerative diseases such as muscle dystrophy, multiple sclerosis, Parkinson's disease, amyotrophic lateral sclerosis, Alzheimer disease, sepsis, malignant and non-malignant tumors; as well as chemical-, heat- and irradiation-induced inflammatory responses.
  • diseases e.g., psoriasis, arthritis, Crohn's disease, degenerative diseases such as muscle dystrophy, multiple sclerosis, Parkinson's disease, amyotrophic lateral sclerosis, Alzheimer disease, sepsis, malignant and non-malignant tumors; as well as chemical-, heat- and irradiation-induced inflammatory responses.
  • Fibrinopeptide A is known to have anti-inflammatory properties (Ruhenstroth- Bauer G et al., Inflammation 5, 343-351, 1981) but its combination with histone H2A fragment, anti TNF antibodies and anti IL1 antibodies, resulting in significant anti- inflammatory, is novel.
  • the histone H2A peptide fragment having residue numbers 36-44 is novel, and its anti-inflammatory properties are also novel. Smaller fragments of these peptides or other analogs are also claimed.
  • fibrinopeptide A had weak anti-inflammatory effect, however, its combination with the histone H2A fragment and anti TNF antibodies and anti ELI antibodies, gives a novel synergistic effect.
  • Protection can be achieved by prophylactic treatment, i.e. treatment with the factor (5 minutes to 3 days) prior to the noxious stimuli protects the individual against bums. Protection can also be achieved by post-exposure treatment with the factor.
  • the dose for administration will be determined in accordance with the condition to be treated.
  • Therapeutic doses are defined as the range of doses able to induce a statistically significant reduction in damage compared with that suffered by an individual not receiving treatment with the protective factor. Protection against noxious stimuli may also manifest itself in terms of decreased toxicity as expressed by secondary criteria, as are well known in the art, including reduction in weight loss suffered after exposure to noxious chemicals, or in terms of maintenance of food consumption.
  • tissue protection will also be influenced by the general health conditions of the subject to be treated, which will relate to various factors including but not limited to the pre-existence of any underlying pathology, as well age and other factors.
  • the route of administration will depend on the condition that it is intended to treat with the factor. Suitable routes of administration include but are not limited to parenteral injections, e.g., intradermal, intravenous, intramuscular, intralesional, subcutaneous, intrathecal, and any other mode of injection as is known in the art.
  • compositions of the invention relate to any pharmaceutically acceptable formulation of the factors of the invention comprising at least one diluent or excipient, as is well known in the art.
  • This term is understood to encompass aqueous solutions, cosolvent solutions, dry powders for solution, solid formulations, formulations for inhalation, suppositories, sustained release formulations, gels, lotions, creams, emulsions, liposomes and the like.
  • the formulation may further comprise isotonicity agents, preservatives, buffers cryoprotectants and the like as is well known in the art. It is further understood that the factors of the invention may be formulated or administered together with additional active ingredients as may be required to treat the condition of the patient.
  • modified peptide or “peptide analog” may be used herein interchangeably, and relate to a peptide that undergoes chemical modification for stabilization against proteolytic and other degradative activities and/or physical or chemical activities.
  • Modified peptides include, but are not limited to, N-methylation of one or several peptide bonds, retro-inverso modification of one or several peptide bonds, substitutions of amino acids by conservative replacements such as are known in the art, addition or omission of amino-terminal or carboxy-terminal amino acids.
  • introduction of proline to the amino terminus may confer resistance against aminopeptidases
  • amidation of the carboxy terminus to produce carboxamide may confer resistance against carboxypeptidases.
  • Tissues that can be protected extends to all kinds of tissues or cells in the body including skin and all skin layers, subcutaneous tissues, muscles, mucosal membranes, neuronal tissue, lungs, upper and lower airways, eyes, gastrointestinal tract, urinary tract, bones and bone marrow.
  • the protective factor can be used for accelerated healing of and prevention of development of wounds including decubitus, ulcers (also induced by drugs), internal and external wounds, abscesses and various bleedings.
  • the protective factors may be used for protection of bone marrow, intestinal epithelium, hair follicles and other sensitive tissues against chemotherapeutic agents such as anticancer drugs, immunosuppresive agents, irradiation and other noxious stimuli.
  • the protective factors may be used for treatment and protection on the central and peripheral nervous systems against noxious stimuli caused by, but not limited to, chemicals, drugs, all kinds of irradiation and mechanical stress.
  • the factors may also serve in treatment of a variety of mental diseases and mental-related syndromes.
  • the protective factors may also be used for treatment or prevention of tissue damage including, but not limited to, neuronal, neurological, hepatic, nephrologic, urologic, cardiac, pulmonary, gastrointestinal, visual, audiologic, spleen, bone, bone marrow and muscular defects.
  • Treatment or prevention of tissue damage may be accomplished in the fetus, newborn, child, adolescent as well as in adults and old persons, whether the condition or disorder to be treated is spontaneous, of traumatic etiology, as a congenital defect or as a teratogenic phenomenon.
  • the protective factors may be used for treatment or prevention of diseases or syndromes related to, or involving inflammatory processes or inflammatory mediators.
  • the term "protective factors” relates to the materials, which elute under the specific chromatography conditions exemplified hereinbelow at 22-27 min retention time.
  • the currently preferred factors are peptides ITI, TV, VI, VII. and all of their analogs, variants and derivatives.
  • Protective factors relates to all of these peptides each as a single compound and/or all possible combinations of two or more of these peptides, including modified peptides.
  • the well was then attached to the animal back, and tested to ensure that liquid inside the well did not leak out.
  • the skin area covered by the well was 1.13 sq. cm.
  • Eight such wells were constructed on the back of each guinea pig. Maintenance of anesthesia during the experiment was achieved by 15mg/kg pentobarbital ip. Each area inside the well was exposed, for 5 seconds, to the base (1.2 cm diameter) of a glass beaker (4.5 cm height) containing 90°C water.
  • Topical application of the fresh liquid obtained from squeezing cut leaf of Aloe vera was shown to protect guinea pigs against thermal bums.
  • Aloe Vera liquid instead the povidone-iodine or iodine ointment as a protectant.
  • Aloe Vera liquid obtained by squeezing cut Aloe Vera leaf was applied to cover the entire area of the skin.
  • the Aloe Vera was replaced by 10% povidone iodine solution or by 2% iodine tincture.
  • the anesthetized animal was exposed to a preheated (75°C) metal (3x5 cm) for 5sec. Then iodine tincture was applied every 10-15 minutes (to keep continuous contact with iodine solution). After two hours the liquid was washed out with water, and the skin was gently dried with a filter paper.
  • the animal was sacrificed, the treated skin was removed from the animal, put on a glass petri dish (9 cm diameter), and sliced into pieces of about 2x2x2 mm.
  • the slices were extracted with 40 ml ethanol containing 800 ⁇ l acetic acid in a 260 ml plastic tissue culture flask incubated vertically at 37°C for 1 hour and mixed with magnetic stirrer. Later on it was found that incubation at 4°C for 1 hour might give better results.
  • the extract was evaporated by rotary evaporator and Speed Vac to a final volume of lOO ⁇ l.
  • the extract was diluted with 2% acetonitril/0.08% trifluoroacetic acid in double distilled water (solvent A of high performance liquid chromatography (HPLC) gradient system).
  • the tubes were centrifuged (7 min X 15,000g), filtered with 2 ⁇ m filter, and subjected (50 ⁇ l) to HPLC (Applied Biosystem equipped with 140 B solvent delivery system and 1000 S diode array detector) using Vydac C18 column (2.1X250 mm) and C18 precolumn.
  • Peptide ITI was found to be a partial sequence of Histone H2A that its carboxy terminal Gly was substituted by Ala.
  • Peptide VI was identified as guinea pig fibrinopeptide A, while peptides TV and VII differ from peptide VI by omission and addition, respectively, of the amino terminal threonine moiety.
  • Example 6 shows 10.9% increase in body weight in peptide combined with proteinase inhibitors-treated animals while treatment with proteinase inhibitors only increased body weight by 6.8%.
  • iodine (1 ml, 2% in tetraglycol) was applied into the well for 2 hours. In the end of procedure the iodine was sucked out and the treated skin sites of 4 guinea pigs were removed, cut into small pieces and extracted in 100 ml ethyl alcohol in ultrasonic bath at 37°C for 1 hour.
  • the skin extract was concentrated to a volume of 20-40 ⁇ l, diluted in 800 ⁇ l saline (0.9% NaCl), and injected intradermally into guinea pigs.
  • Each guinea pig received 4 injections (50 ⁇ l each at 4 sites, two at each side of the back) 5 min before the heat stimulus (75°C water for 10 seconds) by the well system.
  • Additional control guinea pigs were injected with saline, and exposed to heat in the same manner.
  • the animals were daily checked for gross pathology. Ulceration area was measured by digital camera equipped with a ruler.
  • Results are expressed as mean ⁇ SEM of 18 extract-injected sites and 18 saline-injected sites (control) using the Mann- Whitney (two-tailed) for statistical evaluation of the differences between the groups. *P ⁇ 0.05 As demonstrated in Fig. 1 , intradermal injection of a skin extract into guinea pig skin 5 min before the heat stimulus reduced the ulceration area by 69% as compared to the control animals (injected with saline only).
  • Example 2 Protective effect of isolated HPLC fraction Intradermal injection of the factor (purified by HPLC from the skin extract) into guinea pigs that were exposed to heat (75°C for 20 sec on a circle of 0.5 cm diameter), or to chemical (500 ⁇ g/0.7x0.7 cm mechlorefhamine hydrochloride) stimuli was evaluated for gross pathology after 3 days. The injection sites were located 1 cm from the edge of the application site. A control guinea pig was injected with an HPLC fraction eluted at the same retention time (25 min), but without the factor.
  • Table 1 HPLC fraction-induced protection against thermal and chemical stimulation in the guinea pig skin model.
  • each well was exposed to 1 ml 75°C distilled water for 10 sec, then water was sucked out and well was removed.
  • Each injection contained the following components:
  • Hvdrophobic proteinase inhibitors Pepstatin A and Chymostatin, 1 mg of each inhibitor was dissolved in 1ml dimethylsulfoxide (DMSO);
  • mice were daily checked for gross pathology, Ulceration area was measured by digital camera with the aid of a ruler. Results are expressed as mean ⁇ SEM (each column represents 12 exposure sites) using the Mann- Whitney (two- tailed) for statistical evaluation of the differences between the groups.
  • Example 4 Protective effect of peptides against chemical skin burns.
  • each well was exposed to 1 ml 75°C distilled water for 10 sec, then water was sucked out and well was removed.
  • Each injection contained the following components:
  • Example 5 Protective effect of peptides against thermal skin burns.
  • each well was exposed to 1 ml 75°C distilled water for 10 sec, then water was sucked out and well was removed.
  • Each injection contained the following components:
  • H) Peptide solution for injection: 4 tubes containing solution B were prepared. Each tube contained 5mg peptide of one sort, so that each tube contained all kinds of inhibitors and one kind of peptide. Each peptide is indicated by its Roman numeral plus I (inhibitors of proteinases) (n 9). Additional experiment in which a mixture of all four peptides (ffl, IV, VI, VII),
  • Control solution for injection 1ml 0.9% NaCl containing 5 ⁇ l DMSO
  • Example 6 Effect of peptides against body weight loss induced by sulfur mustard and thermal injury.
  • each well was exposed to 1 ml 75°C distilled water for 10 sec, then water was sucked out and well was removed.
  • Each injection contained the following components:
  • Example 7 Protective effect of N-methylated analogs of peptide III against mustard gas-induced skin lesions
  • Percent protection means % of reduction in ulceration area in the peptide-treated group in comparison to the control (vehicle) group.
  • Fig. 7 Protective effect of peptides ffl, 3b and IV against SM in the mouse ear swelling model.
  • mice Male ICR mice ( ⁇ 25g) were anesthetized by pentobarbital sodium 60mg/kg ip (0.1ml/25g BW of 1.5% solution) and placed on their abdominal side. Anesthesia was maintained by 0.03ml/25g BW whenever needed. Peptides ffl, 3b and TV (dissolved in
  • mice received 0.9% NaCl injections. Within 5 min after injection, the outer side of each ear was exposed to 0.317 mg SM (5 ⁇ l of 1 :20 dilution in dichloromethane). Mouse ear thickness was measured 48 hours after exposure using micrometer (Model PK-0505, Mitutoyo Corporation, Japan). Edema was assessed by the difference between ear thickness measured after and prior to exposure. Animals were sacrificed 48 hours after exposure. Results are the mean ⁇ SE using the Mann Whitney U test for statistical evaluation of the difference between Cont and peptide TV group. The asterisk denotes statistical significance at the level of *p ⁇ 0.01.
  • Fig. 8 demonstrates the protective effect of the combinations of anti mouse ILl beta antibodies, anti mouse TNF alpha antibodies, peptide TV and peptide 3ml against sulfur mustard -induced inflammatory response as expressed by mouse ear edema. This combination reduced SM-induced ear edema by 31%. Omission of peptide IV from this combination reduced mouse swelling by 15%. Peptide TV only has no protective effect. Thus, peptide TV had a synergistic effect on the protection of the combination of anti mouse ILl beta antibodies, anti mouse TNF alpha antibodies, and peptide 3ml.
  • ILl beta interleukin 1 beta
  • TNF alpha tumor necrosis factor alpha
  • mice Male ICR mice ( ⁇ 25g) were anesthetized by pentobarbital sodium 60mg/kg ip (0.1ml/25g BW of 1.5% solution) and placed on their abdominal side. Anesthesia was maintained by 0.03ml/25g BW whenever needed. Peptides 3m, 3ml, IV and TV (dissolved in 0.9% NaCl), and anti mouse ILl beta (ILlAb) antibodies and anti mouse TNF alpha antibodies (TNFAb) were tested for their ability to protect against SM- induced toxicity.
  • ILlAb anti mouse ILl beta antibodies and anti mouse TNF alpha antibodies
  • Results are the mean ⁇ SE using the Mann Whitney U test for statistical evaluation of the difference between 0.9% NaCl group and experimental groups.
  • the asterisk denotes statistical significance at the level of p ⁇ 0.04.
  • the same statistical significance was observed for the difference between the group of: 5 ⁇ g/injection anti mouse ILl beta antibodies, lO ⁇ g/injection anti mouse TNF alpha antibodies, lOmg/kg 3ml, and the group of: 5 ⁇ g/injection anti mouse ILl beta antibodies, lO ⁇ g/injection anti mouse TNF alpha antibodies, lOmg/kg IN, lOmg/kg 3ml;
  • the same statistical significance was observed for the difference between the group of: 5 ⁇ g/injection anti mouse ILl beta antibodies, lO ⁇ g/injection anti mouse T ⁇ F alpha antibodies, lOmg/kg IV, and the group of: 5 ⁇ g/injection anti mouse ILl beta antibodies, lO ⁇ g/injection anti mouse T ⁇ F alpha antibodies,
  • a "pharmaceutical composition” refers to a preparation of one or more of the peptides described herein, or physiologically acceptable salts or prodmgs thereof, with other chemical components such as physiologically suitable carriers and excipients.
  • the purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.
  • prodrug refers to an agent, which is converted into an active parent drug in vivo. Prodmgs are often useful because in some instances they may be easier to administer than the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent drug is not. The prodrug may also have improved solubility compared to the parent drug in pharmaceutical compositions.
  • excipient refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a compound.
  • excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.
  • compositions may also include one or more additional active ingredients, such as, but not limited to, conventional anti-migraine agents.
  • Pharmaceutical compositions of the present invention may be manufactured by processes well known in the art, e.g., by means of conventional mixing, dissolving, granulating, grinding, pulverizing, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes.
  • compositions for use in accordance with the present invention thus may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active compounds into preparations which, can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.
  • the compounds of the invention may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological saline buffer.
  • physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological saline buffer.
  • penetrants appropriate to the barrier to be permeated are used in the formulation.
  • penetrants for example DMSO, or polyethylene glycol are generally known in the art.
  • the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers well known in the art.
  • Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for oral ingestion by a patient.
  • Pharmacological preparations for oral use can be made using a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries if desired, to obtain tablets or dragee cores.
  • Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carbomefhylcellulose; and/or physiologically acceptable polymers such as polyvinylpyrrolidone (PVP).
  • disintegrating agents may be added, such as cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
  • enterocoating are useful as it is desirable to prevent exposure of the peptides of the invention to the gastric environment.
  • Dragee cores are provided with suitable coatings.
  • suitable coatings may be used which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures.
  • Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
  • compositions which can be used orally, include push-fit capsules made of gelatin as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol.
  • the push-fit capsules may contain the active ingredients in admixture with filler such as lactose, binders such as starches, lubricants such as talc or magnesium stearate and, optionally, stabilizers.
  • the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • stabilizers may be added. All formulations for oral administration should be in dosages suitable for the chosen route of administration.
  • the compositions may take the form of tablets or lozenges formulated in conventional manner.
  • the peptides for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or a nebulizer with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichloro- tetrafluoroethane or carbon dioxide.
  • a suitable propellant e.g., dichlorodifluoromethane, trichlorofluoromethane, dichloro- tetrafluoroethane or carbon dioxide.
  • the dosage unit may be determined by providing a valve to deliver a metered amount.
  • Capsules and cartridges of, e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the peptide and a suitable powder base such as lactose or starch.
  • compositions for parenteral administration include aqueous solutions of the active ingredients in water-soluble form. Additionally, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acids esters such as ethyl oleate, triglycerides or liposomes. Aqueous injection suspensions may contain substances, which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol or dextran. Optionally, the suspension may also contain suitable stabilizers or agents, which increase the solubility of the compounds, to allow for the preparation of highly concentrated solutions.
  • the active ingredient may be in powder form for reconstitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.
  • a suitable vehicle e.g., sterile, pyrogen-free water
  • the compounds of the present invention may also be formulated in rectal compositions such as suppositories or retention enemas, using, e.g., conventional suppository bases such as cocoa butter or other glycerides.
  • compositions herein described may also comprise suitable solid of gel phase carriers or excipients.
  • suitable solid of gel phase carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin and polymers such as polyethylene glycols.
  • compositions suitable for use in context of the present invention include compositions wherein the active ingredients are contained in an amount effective to achieve the intended purpose. More specifically, a therapeutically effective amount means an amount of a compound effective to prevent, alleviate or ameliorate symptoms of a disease of the subject being treated.
  • Toxicity and therapeutic efficacy of the peptides described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the IC ⁇ Q (the concentration which provides 50% inhibition) and the LD50 (lethal dose causing death in 50 % of the tested animals) for a subject compound.
  • the data obtained from these cell culture assays and animal studies can be used in formulating a range of dosage for use in human.
  • the dosage may vary depending upon the dosage form employed and the route of administration utilized.
  • the exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition. (See e.g., Fingl, et al., 1975, in "The Pharmacological Basis of Therapeutics", Ch. 1 p.l).
  • dosing can also be a single administration of a slow release composition, with course of treatment lasting from several days to several weeks or until cure is effected or diminution of the disease state is achieved.
  • compositions to be administered will, of course, be dependent on the subject being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, and all other relevant factors.
  • the following example is an illustration only of a method of treating a subject with a peptide according to the invention, in order to treat a pathological condition associated with tissue trauma or a related condition, and is not intended to be limiting.
  • the method includes the step of administering the protective peptide, in a pharmaceutically acceptable carrier as described above, to a subject to be treated.
  • the medicament is administered according to an effective dosing methodology, preferably until a predefined endpoint is reached, such as a reduction or amelioration of the pathological condition in the subject.
  • the protective factors may be used for treatment and protection on the central and peripheral nervous systems against noxious stimuli caused by, but not limited to, chemicals, drugs, all kinds of irradiation and mechanical stress.

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Abstract

Selon l'invention, des facteurs peptidiques isolés d'une peau ayant subi une lésion chimique ou thermique et le traitement au moyen de préparations d'iodine peuvent réduire ou soigner l'extension de la blessure lorsque respectivement on les administre ou on les applique à d'autres animaux. Des peptides spécifiques, notamment des fragments d'histone H2A et de fibrinopeptide A, et des dérivés de ces peptides utilisés de préférence sont décrits, ainsi que des compositions pharmaceutiques et des procédés d'utilisation de ces peptides. Selon l'invention, les extraits, les peptides et les compositions pharmaceutiques sont utiles dans la prévention et dans le traitement d'états pathologiques inflammatoires et d'une exposition à des stimuli nocifs.
PCT/IL2002/000713 2001-08-29 2002-08-29 Facteurs de protection contre des inflammations, des brulures et des stimuli nocifs WO2003017920A2 (fr)

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AU2002329017A AU2002329017A1 (en) 2001-08-29 2002-08-29 Protective factors against inflammation, burns and noxious stimuli
EP02765303A EP1572069A4 (fr) 2001-08-29 2002-08-29 Facteurs de protection contre des inflammations, des brulures et des stimuli nocifs
US10/790,888 US7238656B2 (en) 2001-08-29 2004-03-01 Protective factors against inflammation, burns and noxious stimuli
US11/750,408 US7605132B2 (en) 2001-08-29 2007-05-18 Protective factors against inflammation, burns and noxious stimuli

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IL145181 2001-08-29
IL145181A IL145181A0 (en) 2001-08-29 2001-08-29 Protective factors against burns and noxious stimuli

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US11/527,162 Continuation-In-Part US7528227B2 (en) 2001-08-29 2006-09-25 Histone H2A peptide derivatives and uses thereof

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WO2005090387A2 (fr) * 2004-03-23 2005-09-29 Yissum Research Development Company Of The Hebrew University Of Jerusalem Derives de peptide d'histone h2a et analogues, et methodes d'utilisation
WO2006101187A1 (fr) * 2005-03-22 2006-09-28 Rohto Pharmaceutical Co., Ltd. Peptide qui augmente la production de collagene ou d’acide hyaluronique
US7528227B2 (en) 2004-03-23 2009-05-05 Yissum Research Development Company Of The Hebrew University Of Jerusalem Histone H2A peptide derivatives and uses thereof
US7605132B2 (en) 2001-08-29 2009-10-20 Yissum Research Development Company Of The Hebrew University Of Jerusalem Protective factors against inflammation, burns and noxious stimuli
WO2011128884A1 (fr) 2010-04-11 2011-10-20 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Extrait et peptides dérivés du groupe oryza sativa japonica et leurs utilisations
CN106397575A (zh) * 2016-10-20 2017-02-15 上海懿贝瑞生物医药科技有限公司 一种具有胞外组蛋白毒性抑制作用的多肽制备方法及用途

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WO2000064466A1 (fr) * 1999-04-28 2000-11-02 Northwestern University Localisation d'autoepitopes de peptides majeurs pour lymphocytes t nucleosomes specifiques de lupus erythemateux aigu dissemine
CN1296974A (zh) * 1999-11-23 2001-05-30 上海博容基因开发有限公司 一种新的多肽——人组蛋白h2a.21和编码这种多肽的多核苷酸

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DATABASE MEDLINE [Online] ELGJO K. ET AL.: 'Proliferation-dependent effect of skin extracts (chalone) on mouse epidermal cell flux at the G1-S, S-G2 and G2-M transitions', XP008099665 Retrieved from STN Database accession no. (83199201) & VIRCHOWS ARCHIV. B vol. 42, no. 2, 1983, pages 143 - 151 *
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Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7605132B2 (en) 2001-08-29 2009-10-20 Yissum Research Development Company Of The Hebrew University Of Jerusalem Protective factors against inflammation, burns and noxious stimuli
WO2005090387A2 (fr) * 2004-03-23 2005-09-29 Yissum Research Development Company Of The Hebrew University Of Jerusalem Derives de peptide d'histone h2a et analogues, et methodes d'utilisation
WO2005090387A3 (fr) * 2004-03-23 2006-02-02 Univ Jerusalem Yissum Res Dev Derives de peptide d'histone h2a et analogues, et methodes d'utilisation
US7528227B2 (en) 2004-03-23 2009-05-05 Yissum Research Development Company Of The Hebrew University Of Jerusalem Histone H2A peptide derivatives and uses thereof
WO2006101187A1 (fr) * 2005-03-22 2006-09-28 Rohto Pharmaceutical Co., Ltd. Peptide qui augmente la production de collagene ou d’acide hyaluronique
US7989590B2 (en) 2005-03-22 2011-08-02 Rohto Pharmaceutical Co., Ltd Peptides that increase collagen or hyaluronic acid production
WO2011128884A1 (fr) 2010-04-11 2011-10-20 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Extrait et peptides dérivés du groupe oryza sativa japonica et leurs utilisations
EP2558110A1 (fr) * 2010-04-11 2013-02-20 Yissum Research Development Company of the Hebrew University of Jerusalem Ltd. Extrait et peptides dérivés du groupe oryza sativa japonica et leurs utilisations
EP2558110A4 (fr) * 2010-04-11 2013-09-25 Yissum Res Dev Co Extrait et peptides dérivés du groupe oryza sativa japonica et leurs utilisations
US8765679B2 (en) 2010-04-11 2014-07-01 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Extract and peptides derived from Oryza sativa Japonica Group and uses thereof
CN106397575A (zh) * 2016-10-20 2017-02-15 上海懿贝瑞生物医药科技有限公司 一种具有胞外组蛋白毒性抑制作用的多肽制备方法及用途

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WO2003017920A3 (fr) 2006-08-10
IL145181A0 (en) 2009-02-11
EP1572069A2 (fr) 2005-09-14
EP1572069A4 (fr) 2009-06-24
AU2002329017A8 (en) 2006-11-09
AU2002329017A1 (en) 2003-03-10

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