Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • C07K14/665—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin
• • 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
  • A61P25/30—Drugs for disorders of the nervous system for treating abuse or dependence
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K17/00—Carrier-bound or immobilised peptides; Preparation thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides

Definitions

• the present invention generally relates to dynorphin analogues, and more particularly to novel dynorphin analogues that can be used with narcotic analgesics, such as opiate alkaloids.
• narcotic analgesics such as opiate alkaloids.
• the endogenous opioids exist in multiple forms in the central nervous system and include the dynorphins, which are a series of peptides derived from the precursor prodynorphin (proenkephalin B) .
• the first of the dynorphins to be isolated was the 17 amino acid peptide having the structure shown (and designated SEQ ID N0:1) , sometimes also referred to as "dynorphin A (1- 17)":
• dynorphins Unlike either the enkephalins or the endorphins, many of the dynorphins interact with high affinity with all three major opioid receptor types ( ⁇ , S , and K) .
• the dynorphins are also nearly unique among endogenous opioids in that they are not analgesic in the brain, although they may be in the spinal cord.
• dynorphin A (1-13) potentiates the analgesic effect in tolerant hosts.
• Dynorphin was found useful in conjunction with a narcotic analgesic in order to reduce the amount of narcotic analgesic administered per dose.
• AA8 i.s i.soleucm. e, leucme, or lys e
• AA9 is arginine or proline
• AA is proline
• a carbonyl carbon at the AA terminus is amidated.
• dynorphin (1-10) amide analogs appear to be a more potent and selective analog than dynorphin (1-13).
• SEQ ID NO: 3 represents a particular one of these dynorphin (1-10) amide analogs (where the C-terminal Pro is amidated) :
• dynorphin-related ⁇ pioid peptides such as the SEQ ID NO:2 and SEQ ID NO:3 peptides.
• endogenous opioid peptides condition the sensitivity of the peripheral nerves to stimuli that affect heart rate and blood pressure.
• circulating opioid peptides under normal conditions, are operating to control the sensitivity of these peripheral sites of the autonomic nervous system to such endogenous substances.
• Use of the dynorphin- related peptides in treating high blood pressure appears to modify the autonomic nervous system so as to amplify and maintain the intensity of endogenous opioid peptides.
• a mode of action may be by increasing the sensitivity of visceral afferent receptors.
• Enkephalin analogues that are conformationally constrained by a cyclic structure (such as with a disulfide bridge) are described by U.S. Patent 4,518,711, issued May 21, 1985, inventors Hruby et al. These compounds are said to have increased rigidity and increased delta receptor specificity if the half- cysteine in the 2 position is replaced by half- penicillamine ( ⁇ , ⁇ -dimethyl half-cysteine) .
• dynorphin analogues have become known that have cysteine replacements at the amino acid residue 5 (usually leucine) and at the amino acid residue 11 (usually lysine) .
• the amino acid residue 8 (usually an isoleucine) and the amino acid residue 13 (usually a lysine) have similarly been replaced by cysteines in a bridged relationship.
• the bridges, or cyclic structures, appear to assist in stabilizing the dynorphin analogues against in vivo degradations.
• mice having been treated with chronic morphine were immuno- suppressed, whereas use of either SEQ ID NO:2 or SEQ ID NO:3 was found to block the opioid inhibition of acrophage-colony stimulating factor of morphine in a dose-dependent manner.
• SEQ ID NO:2 or SEQ ID NO:3 was found to block the opioid inhibition of acrophage-colony stimulating factor of morphine in a dose-dependent manner.
• novel peptides are described that have at least six amino acid residues, are analogues of dynorphin, but are des-Tyr with respect to the endogenous dynorphin.
• novel peptides may be formulated in a pharmaceutically acceptable solution or with a pharmaceutically acceptable carrier, and are usefully administered to a host tolerant to a narcotic analgesic in order to potentiate activity of the narcotic analgesic and/or to block withdrawal symptoms.
• Additional uses include the reversal of at least some neurologic deficit in treating cerebral and spinal ischemia, in inhibiting respiratory depression or gastroenteric spasms produced by narcotic analgesics to a naive host, as an adjunct for ' anti- inflammatory medication, and in blocking narcotic induced immune impairment in a host whose immune system has been impaired by a narcotic analgesic.
• novel peptides of the invention generally have similar activity to endogenous dynorphin (SEQ ID N0:1), to dynorphin with thirteen amino acids (SEQ ID NO:2), and to dynorphin in amide form with ten amino acids (SEQ ID NO:3); however, it is surprising that the novel compounds, which are des-Tyr with respect to such previously known dynorphin compounds, exhibit similar biological activity because the N-terminal tyrosine has been considered a substantially universal requirement for recognition of opioid peptides by opioid receptors.
• narcotic analgesic for example an opiate alkaloid such as morphine
• a narcotic such as methadone
• the various, known side effects, such as respiratory depression and constipation, which result from chronic treatment with high doses of narcotics can be lessened by practice of the invention.
• Novel peptides of the invention have at least six amino acids for the various desirable therapeutic applications. Where the at least six amino acid residues are present, then they preferably are in the sequence: Gly-Phe-Leu-Arg-Arg-Ile (SEQ ID N0:22) .
• novel peptides can be viewed as having amino acid residues analogous to endogenous dynorphin (SEQ ID N0:1), but where the novel peptides are des-Tyr, as shown by SEQ ID NO:1
• Gly-Gly-Phe-Leu-Arg-Arg-Ile-Arg-Pro-Lys-Leu-Lys- Trp-Asp-Asn SEQ ID NO:4
• Gly-Gly-Phe-Leu-Arg-Arg-Ile-Arg-Pro-Lys-Leu-Lys- Trp-Asp SEQ ID N0:5
• Trp SEQ ID NO: 6
• Gly-Gly-Phe-Leu-Arg-Arg-Ile-Arg-Pro-Lys-Leu-Lys SEQ ID NO:7 ;
• any one or two of the residues may be replaced with the same or a different amino acid residue in the D- configuration (to increase in vivo stability) , such as where the N-terminal Gly is replaced by D-Ala, or a modification for conformational stability or rigidity may be made, such as where a plurality of the specified amino acid residue are replaced by moieties capable of forming a cyclic structure, or bridge (e.g., the disulfide bridge) .
• Novel peptides for this invention can also be viewed as des-Tyr-Gly, as shown by SEQ ID NOS:13-22, which can similarly be modified to increase in vivo stability and conformational stability as already described for SEQ ID NOS:4-12:
• novel peptides illustrated by SEQ ID NOS:l-22 can be in the acid or amide form at the C- terminus. Further, the usual isoleucine amino acid residue in SEQ ID NOS:4-22 can be replaced with leucine or lysine, and the usual arginine at position 8 of SEQ ID NOS:4-ll and at position 7 of SEQ ID NOS:13-21 can be replaced with proline.
• One model potentially useful for choosing particular modifications is based upon a prediction that receptor selectivity of opioid peptides is governed by their net charge and/or amphiphilic moment, in addition to the structural and conformational requirement ' s of a particular opioid receptor type ( ⁇ , ⁇ S, and K ) .
• This model predicts that opioid peptides with a net positive charge would show ⁇ -receptor preference while neutral and negatively charged opioid peptides would preferentially interact with the cS-receptor, and the model has been used, for example, in designing small peptides by Schiller et al., J. of Med. Chem. , 32 , pp. 698-703 (1989).
• Novel peptides of the invention are preferably formulated in a pharmaceutically acceptable solution or with a pharmaceutically acceptable carrier, and then are administered in conjunction with a narcotic analgesic.
• the peptide may b-e formulated with a wide variety of physiologically acceptable carriers, such as aqueous saline and phosphate buffered saline, and may include physiologically acceptable excipients, such as glucose, mannitol, or the like.
• dynorphin (1- 17) also known as SEQ ID N0:1, but when des-Tyr (and thus described as SEQ ID NO:23), retains sufficient of the desired activity in conjunction with narcotic analgesics to be useful.
• the present invention is useful with sub ⁇ stantially all narcotic analgesics, and more preferably the opiate alkaloids and opioid peptides (both synthetic and natural) .
• the invention is useful with the various alkaloids of opium such as morphine, morphine salts (such as morphine hydrobromide, morphine hydrochloride, morphine muscate, morphine oleate, morphine N-oxide, and morphine sulfate) , and morphine analogs such as normorphine, diacetyldihydromorphine, diacetylmorphine hydrochloride, codeine, and diacetylmorphine (heroin) .
• morphine salts such as morphine hydrobromide, morphine hydrochloride, morphine muscate, morphine oleate, morphine N-oxide, and morphine sulfate
• morphine analogs such as normorphine,
• narcotic analgesics with which the present invention may be used include alphaprodine, methadone, meperidine, levorphanol, propoxyphene, fentanyl and its analogues, oxymorphone, anileridine, dilaudid, and metopon. Uses can be extended to the peptide analgesics, such as enkephalins and ⁇ -endorphin analogs.
• narcotic analgesics As is well known, continued use of these narcotic analgesics leads to habituation or addiction, and use of one leads to cross-tolerance for the others. However, despite their abuse potential, these narcotic analgesics have therapeutic uses, for example with patients requiring chronic treatment to ease pain.
• narcotic analgesics can be, and are, studied in various mammalian species besides humans, since practical and governmental considerations frequently require that studies be first done in small rodents and/or monkeys before the analgesic properties of pharmaceuticals are tested with humans.
• drugs that have morphine-like properties in mammals other than humans have been found to be morphine-like in humans, and a variety of analgesic assays have been developed with animals which have gained widespread acceptance for predicting properties in humans.
• the present invention includes administering a dose of one of the analogues SEQ ID NOS:4-23, or a modified version thereof as has been described, to a host in conjunction with administering a dose of a narcotic analgesic, wherein the administration is within at least about 30 minutes of the narcotic analgesic dose.
• the administering is by administering a single, admixed dose where the narcotic analgesic, is morphine, a morphine analogue, or a morphine salt, or other peptide analgesics.
• narcotic analgesic is morphine and is to a naive patient
• a normal dosage is on the order of about 5 mg i.v., assuming a body weight of about 70 kg. It is believed a suitable dose of the dynorphin analogue, administered in conjunction with the analgesic, is from about 30-1500 ⁇ g per kg body weight.
• the dynorphin analogue does not potentiate the narcotic analgesic in an initially naive host, as the patient continues in an extended treatment with narcotics to ease pain, the amount of narcotic required to produce a sufficient level of analgesia over the treatment period will be less than without use of dynorphin analogue in conjunction with the narcotic. As a consequence, the various undesirable side effects of repeated, high doses of narcotics, can be lessened.
• the dosage in tolerant patients may be determined as follows. A first, or sufficient, dose of the narcotic analgesic is determined which would be sufficient to produce analgesia in the host. However, instead of administering the sufficient dose, a predetermined dose of the narcotic analgesic is administered. This predetermined, or second, dose includes less of the narcotic analgesic than would be sufficient to produce analgesia in the host. The actually administered dose of narcotic analgesic is supplemented with dynorphin analogue.
• the supple ⁇ mentation is preferably sufficient to produce a level of analgesia in the host which is substantially equivalent to the level of analgesia were solely the narcotic analgesic to have been administered.
• the first or sufficient dose, the lower, second dose, and the supplementing dose will vary depending upon the patient's particular level of tolerance to the narcotic analgesic, and will normally be determined by the treatment physician.
• Another therapeutic method of use is in treating addicts to substantially block withdrawal symptoms.
• methadone usually methadone hydrochloride
• clonidine another drug, such as clonidine
• methadone is itself addictive, and clonidine is believed to simply mask withdrawal symptoms.
• dynorphin analogues block the withdrawal symptoms of morphine addicted hosts, yet are at least 100 times less addictive than morphine.
• the administrating of the present invention may be used to assist in blocking withdrawal symptoms in therapeutic treatments of narcotic addicts being treated for addiction.
• administering a dose of dynorphin analogue to a host tolerant to narcotic analgesics will provide a significantly more desirable treatment in treating narcotic addiction.
• Novel peptides of the invention can further be used partially to reverse neurologic deficits in cerebral ischemia. It is believed that factors affecting response to therapy for cerebral ischemia in accordance with the present invention include the dosage, the route of administration, and duration of therapy. However, blood pressure does not appear to be a factor affecting response to therapy for cerebral ischemia in accordance with the present invention.
• therapy is initiated by administering a dose of the dynorphin analogue and then preferably continued by administering subsequent doses.
• the initial dose may be from about 1.0 ⁇ g/kg of patient's weight to about 10 g/kg of patient's weight, more preferably about 100 ⁇ g/kg of patient' ;s weight, and can be delivered by various means known to the art, such as by intravenous injection ("I.V.”).
• Subsequent doses may also be delivered by various means known to the rt, such as by injections or through topical applications in conjunction with a drug carrier, such as dimethyl sulfoxide.
• a drug carrier such as dimethyl sulfoxide.
• continuous infusion may be by use of an implanted mini-pump, or by I.V.
• the doses may be gradually reduced, or titrated.
• Analgesia was measured by the tail-flick method of D'Amour and Smith, J. Pharmac. Exp. Ther. , 72 , pp. 74-79 (1941) , incorporated herein by reference, as modified by Tulunay and Take ori, J. Pharmac. Exp. Ther. , 190 , pp. 395-400 (1974), incorporated herein by reference.
• ED 50 e.g., effective does for 50% of the test group
• the animals' responses were made quantal by establishing an endpoint which represented a significant increase in reaction time.
• the endpoint was an increase in reaction time of an individual animal of greater than 3 SD (e.g., standard deviation) of the control mean reaction time for all animals used in the assay.
• the usual control mean reaction time was 3.1 ⁇ 0.05 sec.
• Nonresponding animals were removed from the heat stimulus when reaction time exceeded 10 sec. to avoid tail damage.
• Drugs were injected 30 minutes prior to testing, unless otherwise indicated. Morphine was injected subcutaneously (s.c.) whereas the peptides were injected (i.v.. in 4 ml saline.
• Morphine tolerance was established by implanting morphine pellets, 75 mg base, subcutaneously by the method of Way et al., J. Pharmac. Exp. Ther. , 167, pp. 1-8 (1969) , incorporated herein by reference.
• the drug used was morphine sulfate (Mallinckrodt Chemical Works, St. Louis, MO) .
• the ED 50 values, their 95% confidence limits and significance of the potency ratio between two ED 50 values were determined by the method of Litchfield and Wilcoxon, J. Pharmac. Exp. Ther. , 96 , 99-113 (1949), incorporated herein by reference.
• the morphine tolerant (addicted) animals had pellets implanted for three days. The animals were then challenged with naloxone doses (while the morphine pellet remained in place) . The naloxone places the animals into a state of narcotic withdrawal and the animal exhibits withdrawal symptoms because naloxone is an antagonist of morphine.
• Table 1 summarizes the data for the control animals and for groups of animals treated with three different peptides. Each peptide was in a dose of 5 ⁇ ol/kg i.v. before administration of the naloxone.
• the dynorphin (3-13), SEQ ID NO:17 is also a novel compound.
• this novel compound is des-Tyr-Gly, it has a potency ratio of 2. That is, addicted animals that were pretreated with this novel peptide before receiving the narcotic antagonist were only half as dependent on morphine as addicted animals which were not so pretreated.
• the cyclic dynorphin amide compound used (where the normal leucine at the 5 position and the normal lysine at the 11 position had been replaced by cysteines, whose disulfide bridge provides conformational stability) had a potency ratio of 2.6, while the des-Tyr compound surprisingly had a potency ratio of 3.7.
• the antinociceptive activity ("pain killing") of morphine was tested in morphine tolerant (that is, addicted) mice, as well as in naive, unaddicted (that is, normal) animals. Two base lines were thus established for the two different control animals.
• the first control (unaddicted) animals had a morphine ED 50 of 6.5. This means that when the naive animals were administered 6.5 ⁇ mol/kg s.c. morphine before the tail flick test, then half of those animals felt no pain.
• the second control group which were morphine addicted animals, required an amount of morphine increased by a factor of 8.5 in order for half of the animals to feel no pain.
• the similarly addicted animals when pretreated with the des-Tyr compound (at 2.5 ⁇ mol/kg i.v. 5 minutes before testing) had a significantly decreased amount of morphine necessary for the pain relief.

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• 1993
  • 1993-06-01 AU AU43990/93A patent/AU4399093A/en not_active Abandoned
  • 1993-06-01 JP JP6501537A patent/JPH08501075A/ja active Pending
  • 1993-06-01 CA CA002137916A patent/CA2137916A1/en not_active Abandoned
  • 1993-06-01 CZ CZ943131A patent/CZ281630B6/cs unknown
  • 1993-06-01 EP EP93914274A patent/EP0652765A4/en not_active Withdrawn
  • 1993-06-01 HU HU9403563A patent/HUT70160A/hu unknown
  • 1993-06-01 WO PCT/US1993/005161 patent/WO1993025217A1/en not_active Application Discontinuation
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• 1994
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