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WO1993010784A1 - Composition et procede therapeutiques de prevention de lesions occasionnees par perfusion - Google Patents

Composition et procede therapeutiques de prevention de lesions occasionnees par perfusion Download PDF

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Publication number
WO1993010784A1
WO1993010784A1 PCT/US1992/010116 US9210116W WO9310784A1 WO 1993010784 A1 WO1993010784 A1 WO 1993010784A1 US 9210116 W US9210116 W US 9210116W WO 9310784 A1 WO9310784 A1 WO 9310784A1
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WO
WIPO (PCT)
Prior art keywords
riboflavin
oxygen
composition
tissues
living
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.)
Ceased
Application number
PCT/US1992/010116
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English (en)
Inventor
Donald E. Hultquist
Marshal Shlafer
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.)
University of Michigan Ann Arbor
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University of Michigan Ann Arbor
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Filing date
Publication date
Application filed by University of Michigan Ann Arbor filed Critical University of Michigan Ann Arbor
Publication of WO1993010784A1 publication Critical patent/WO1993010784A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A61K31/525—Isoalloxazines, e.g. riboflavins, vitamin B2

Definitions

  • This invention relates generally to a therapeutic composition and method for protecting human cells, tissues, and organs from reperfusion injury following ischemia, or mitigating against the harmful effects thereof, and more particularly, to a safe and effective pharmacologic intervention procedure comprising administration of a therapeutic composition containing an antioxidant compound, .such as riboflavin or a riboflavin analog, or in alterna ⁇ tive embodiments, pyrroloquinoline quinone.
  • an antioxidant compound .
  • Ischemia the result of reduction of blood flow, and hence, of oxygen to living tissues, is a common clinical event. It is often associated with morbidity and mortali ⁇ ty. Ischemia occurs spontaneously, for example, during cardiac arrest, myocardial infarction, stroke, shock, suffocation, drowning, and acute lung injury.
  • organ e.g., heart, liver, kidney, lung
  • ischemia is induced in the transplanted organ in the period of time between harvesting from a donor and implantation in the recipient.
  • reperfusion injury an additional component of damage to the organs, called reperfusion injury, occurs when oxygen is restored after deprivation of oxygen and blood flow. Reperfusion injury is also attendant to a variety of surgical proce ⁇ dures such as cardiac or vascular surgery.
  • drugs that are called calcium channel blockers drugs that are called calcium channel blockers, agents that allegedly inhibit the breakdown of metabolic fuels, and redpx enzymes such as superoxide dismutase.
  • an object of this invention to provide a pharmacologic intervention to prevent and/or mitigate reperfusion injury to cells, tissues, and organs, after ischemia.
  • the foregoing and other objects are addressed by this invention which provides a composition for treating blood and oxygen-deprived cells, tissues or organs of a living being to prevent reperfusion injury upon reintroduction of blood flow and/or oxygen.
  • the composition comprises an antioxidant compound, preferably in a solution suitable for introduction into living tissues, cells, organs, or organisms, in a therapeutically effective amount.
  • antioxidant compound refers to a compound which, in a biological environment, counteracts the deleterious effects of reactive oxygen species.
  • the antioxidant compound is riboflavin or a riboflavin analog.
  • the antioxidant compound is pyrroloquino- line quinone (PQQ) .
  • the therapeutically effective amount is that amount of antioxidant compound which produces the desired effect, i.e., prevention or mitigation of reperfusion injury. In certain preferred embodiments, about l to 200 micromoles riboflavin in solution has been found to be effective.
  • riboflavin includes riboflavin analogs such as flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetraacetate, riboflavin tetrabutyrate, deriva ⁇ tives of ribitol, and isoalloxazine derivatives, such as proflavin, lumiflavin, lumiflavin-3-acetate, and lumi- chrome.
  • the composition further contains a cellular nutrient.
  • This cellular nutrient may advantageously be glucose.
  • the presence of glucose has been found to reduce the amount of antioxidant compound required for therapeutic effective ⁇ ness. In preferred embodiments, about 5 to 10 millimoles of glucose in the solution has been efficacious.
  • the composition may comprise a sterile, isotonic solution (such as a parenteral fluid) containing about 1 to 200 micromoles of riboflavin and about 5 to 10 millimoles of glucose.
  • a sterile, isotonic solution such as a parenteral fluid
  • the composition may comprise a sterile, isotonic solution containing about 0.1 to 200 micromoles of pyrrolo ⁇ quinoline quinone and about 5 to 10 millimoles of glucose.
  • a living being is treated to prevent reperfusion injury upon reintroduction of oxygen to blood and oxygen-deprived cells, tissues or organs by introducing to the oxygen-deprived cells, tissues or organs of the living being a therapeutically effective amount of an antioxidant compound.
  • the therapeutically effective amount of the antioxidant compound is preferably in the range of about 0.1 to 200 micromoles.
  • a cellular nutrient is co-introduced with the antioxidant compound.
  • the cellular nutrient is preferably glucose which may be present in the range of about 5 to 20 millimolar.
  • the antioxidant compound may be introduced post-reperfusion, it is particularly preferred to introduce the antioxidant compound and/or cellular nutrient, prior to or concurrent with reintroduction of oxygen to the oxygen- deprived cells, tissues or organs of the living being.
  • the antioxi- dant compound and/or glucose can be provided in solution in a therapeutically effective amount and administered by perfusing the cell, tissue, or organ with the solution.
  • a sterile, isotonic solution can be administered systemically to the living being, such as by intravenous injection.
  • the antioxi ⁇ dant compound is coadministered with a cellular nutrient, glucose.
  • Riboflavin also known as vitamin B 2 , is a small molecule which is essential to human nutrition, and hence is naturally present in human tissues in both the intracel- lular and extracellular spaces. Thus, riboflavin would not be antigenic. Moreover, riboflavin is non-toxic and does not appear to have contractile or hemodynamic effects, when administered at low doses. Further, riboflavin passes easily into the cells when administered systemically or directly to the vasculature of an isolated (i.e., trans- plantable) organ.
  • riboflavin refers to riboflavin, riboflavin derivatives (including flavin mononucleotide, flavin adenine dinucleotide, riboflavin tetraacetate, riboflavin tetrabutyrate, and derivatives in which various hydroxyl groups of the ribitol have been derivatized) , and other isoalloxazine derivatives (includ ⁇ ing proflavin, lumiflavin, lumiflavin-3-acetate, lumi- chrome, and derivatives in which chemical groups are substituted on the various carbon and nitrogen atoms of the isoalloxazine ring) .
  • Riboflavin can be obtained commer ⁇ cially from Sigma Chemical Co., St. Louis, MO.
  • the riboflavin is provided in a sterile, preferably isotonic, solution where the concentration of riboflavin in solution would range from about 1 to 200 micromolar.
  • the components of the riboflavin-containing solution should be compatible with living tissue and suitable for administration to living tissue or to a living being. Riboflavin, especially at the concentration herein stated to be efficacious, is compatible with most parenter- al fluids " in which it can be administered. These fluids include typical, well-known intravenous electrolyte- containing and nutrient-containing media, as well as modified solutions, such as those commonly used at the present time by cardiac surgeons in attempts to minimize (with incomplete success) heart damage due to ischemia and reperfusion.
  • the sterile solution also contains a cellular nutrient.
  • the cellular nutrient is glucose in a concentration of approximately 5 to 10 millimolar.
  • Glucose is advantageously coad inistered during riboflavin treat ⁇ ment.
  • the protective effect of riboflavin is a result of the ability of a tissue enzyme to convert it to dihydroriboflavin. Dihydroriboflavin can destroy destruc ⁇ tive oxidants, or reactive oxygen species, which are ' generated in cells during reperfusion of ischemic tissues.
  • the enzyme flavin reductase, catalyzes the reaction between a flavin and NADPH to form dihydroriboflavin and NADP + .
  • This enzyme is present in red blood cells, but has recently been discovered in the cystolic fraction of a number of cells other than red blood cells, specifically bovine liver and rat liver, spleen, kidney, heart, lung, brain, ovary, and testis.
  • Highly reactive compounds, specifically hydroxyl radicals and the Fe(IV)0 and Fe(V)0 oxidation states of hemeproteins, are believed to be responsible for much of the damage during reperfusion injury. It has now been demonstrated that dihydroribo- -flavin efficiently destroys hydroxyl radicals and the higher oxidation states of hemeproteins.
  • riboflavin is believed to permit the ischemic or anaerobic cells to generate greater than normal quantities of dihydroribofla ⁇ vin by the action of cellular flavin reductase on flavin.
  • the dihydroriboflavin would then destroy the destructive agents and protect the cell from reperfusion injury.
  • a cellular nutrient such as glucose
  • PQQ 4,5-dihydro- 4,5-dioxo-lH-pyrrolo[2,3-f]quinolone-2,7,9-tricarboxylic acid, also known as methoxatin
  • PQQ has been shown to be present in a variety of living cells, including mammalian tissues and fluids. We have discovered that PQQ is a high-affinity substrate for the intracellular enzyme flavin reductase. Kinetic studies of this enzyme have revealed that the Michaelis constant, !_ structuring, for PQQ is much lower than the _ tenuus.
  • the enzyme flavin reductase can function more efficiently in cells than with riboflavin.
  • the reduced form of PQQ has been observed to reduce readily the ferric forms of hemeproteins, namely hemoglobin, cytochrome c, and myoglobin.
  • the reduced form of PQQ also reduces the Fe(IV)0, or ferryl form, of myoglobin.
  • PQQ operates in a manner analogous to ribofla- vin to reduce destructive Fe(IV)0 forms of hemeproteins and other reactive oxygen species.
  • PQQ can be purchased as a highly purified compound from Sigma Chemical Company, St. Louis, MO. PQQ is soluble in water and can be provided in a sterile, preferably isotonic, solution where the concentration of PQQ in solution would range from about 0.1 to 200 micromolar.
  • the procedure comprises introducing to the tissue, organ, or systemic circulation, a sufficient amount of antioxidant compound in a sterile, isotonic solution, such that the concentration of the riboflavin in the solution which reaches the ischemic tissues will be in the range of 1 to 200 micromolar, or 0.1 to 200 micromolar for PQQ.
  • the introduction of the antioxidant compound-containing solution is initiated prior to introduction of oxygen.
  • the treatment might begin concurrently with introduction of oxygen.
  • Specific methods of administering the therapeutic composition of the present invention include, without limitation, (a) perfusing the tissue, or organ, in situ ; (b) perfusing an isolated organ or tissue being replaced or transplanted prior to transplantation; or (c) intravenous injection into the patient as part of first line medical intervention, such as during cardiopulmonary resuscitation and advanced cardiac life support techniques.
  • the treat ⁇ ment preferably should continue for a period of time following introduction of oxygen to the ischemic tissues, illustratively from 20 minutes to 60 minutes.
  • riboflavin In riboflavin embodiments where an isolated organ for transplant is being perfused, it is advantageous to use riboflavin rather than an analog of riboflavin, such as riboflavin mononucleotide, which must be metabolized to the flavin.
  • the technique of the present invention is applicable to the protection of tissues in general, however, the most common tissues, or organs, subjected to reperfusion injury are: heart, lung, liver, kidney, intestine, and brain.
  • the riboflavin treatment has been tested on ischemic isolated organs.
  • rabbit hearts were perfused with a physiologic saline solution, the composition of which mimics the cell-free and protein-free components of blood with respect to glucose and salts.
  • the hearts were instrumented in such a way that their function (e.g., ventricular pressure development, heart rates, and perfus- ate flows through the vasculature) could be monitored, and the effluent coming out of the heart could be collected and assayed for chemical markers of cell damage.
  • the hearts were deprived of adequate oxygen for 55 minutes.
  • Some hearts were perfused for an additional 5 minutes with the same oxygen- depriving solution, but containing 20 micromolar ribofla ⁇ vin.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
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Abstract

Une intervention pharmacologique de protection des cellules, tissus et organes humains contre des lésions par reperfusion après ischémie, ou d'atténuation des effets négatifs, consiste à faire passer une composition thérapeutique dans des cellules, des tissus ou des organes privés de sang et/ou d'oxygène, de préférence avant ou en même temps que la réintroduction d'oxygène. Dans un mode de réalisation pratique, la composition thérapeutique est une solution stérile isotonique comprenant environ 1 à 200 micromoles d'un composé anti-oxydant, tel que la riboflavine ou un analogue de riboflavine. Dans d'autres modes de réalisation, ladite composition thérapeutique est une solution stérile isotonique comprenant environ 0,1 à 200 micromoles du composé anti-oxydant, de la pyrroliquinoline quinone. Dans les modes de réalisation préférés, on administre environ 5 à 10 millimoles d'une substance nutritive cellulaire, par exemple du glucose, conjointement avec le composé anti-oxydant.
PCT/US1992/010116 1991-11-25 1992-11-24 Composition et procede therapeutiques de prevention de lesions occasionnees par perfusion Ceased WO1993010784A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US79771091A 1991-11-25 1991-11-25
US07/797,710 1991-11-25
US96095892A 1992-10-14 1992-10-14
US07/960,958 1992-10-14

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WO1993010784A1 true WO1993010784A1 (fr) 1993-06-10

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WO (1) WO1993010784A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2283913A (en) * 1993-10-19 1995-05-24 Radopath Ltd Anti-viral agents comprising flavins
GB2319474A (en) * 1993-10-19 1998-05-27 Radopath Ltd Anti-viral agents
KR100404134B1 (ko) * 2000-01-12 2003-11-05 주식회사 하이폭시 저산소조건에서 동물세포 배양시 세포생존율을 향상시키는방법
WO2003097056A1 (fr) 2002-05-15 2003-11-27 Clf Medical Technology Acceleration Program, Inc. Pyrroloquinoline quinone utilisee pour le traitement des atteintes cardiaques et methodes d'utilisation de cette derniere
EP2011498A4 (fr) * 2006-04-10 2009-11-11 Mitsubishi Gas Chemical Co Agent ameliorant les fonctions cerebrales et aliment fonctionnel contenant l'agent ameliorant
EP3090738A1 (fr) * 2015-05-04 2016-11-09 Universidade de Santiago de Compostela Riboflavine pour le traitement d'un accident ischémique et/ou d'autres maladies associés à l'excitotoxicité du glutamate
JP2018131410A (ja) * 2017-02-15 2018-08-23 ヒノキ新薬株式会社 カスパーゼ−３阻害剤とその用途
US10531655B2 (en) 2011-12-02 2020-01-14 The Regents Of The University Of California Reperfusion protection solution and uses thereof

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0256472A2 (fr) * 1986-08-11 1988-02-24 Lion Corporation Agent cosmétique pour application sur la peau
US4898870A (en) * 1986-08-07 1990-02-06 Sogo Pharmaceutical Company Limited Pyrroloquinoline quinone compounds useful as an enzyme inhibitor
US5080886A (en) * 1990-01-05 1992-01-14 Sterling Drug Inc. Pharmaceutical compositions for the prevention and treatment of oxidant injuries

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4898870A (en) * 1986-08-07 1990-02-06 Sogo Pharmaceutical Company Limited Pyrroloquinoline quinone compounds useful as an enzyme inhibitor
EP0256472A2 (fr) * 1986-08-11 1988-02-24 Lion Corporation Agent cosmétique pour application sur la peau
US5080886A (en) * 1990-01-05 1992-01-14 Sterling Drug Inc. Pharmaceutical compositions for the prevention and treatment of oxidant injuries

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, 04 February 1985, SIDORENKO et al., 102:44801. *
CHEMICAL ABSTRACTS, 08 November 1982, NAGATOMO et al., 97:161422. *
CHEMICAL ABSTRACTS, 20 March 1989, SERFONTEIN et al., 110:101803. *
HARBARD MEDICAL AREA, 08 November 1990, pp. 1-5. *
THE MERCK INDEX, 11th Edition, (1989), citation #7780 at pages 1233-1234 "Rzoflavin". *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2283913A (en) * 1993-10-19 1995-05-24 Radopath Ltd Anti-viral agents comprising flavins
GB2319474A (en) * 1993-10-19 1998-05-27 Radopath Ltd Anti-viral agents
KR100404134B1 (ko) * 2000-01-12 2003-11-05 주식회사 하이폭시 저산소조건에서 동물세포 배양시 세포생존율을 향상시키는방법
WO2003097056A1 (fr) 2002-05-15 2003-11-27 Clf Medical Technology Acceleration Program, Inc. Pyrroloquinoline quinone utilisee pour le traitement des atteintes cardiaques et methodes d'utilisation de cette derniere
JP2005530786A (ja) * 2002-05-15 2005-10-13 シーエルエフ メディカル テクノロジー アクセラレーション プログラム インコーポレイテッド 心損傷の処置のためのピロロキノリンキノンおよびその使用方法
US7276514B2 (en) 2002-05-15 2007-10-02 Charitable Leadership Foundation - Medical Technology Acceleration Program Pyrroloquinoline quinone drugs for treatment of cardiac injury and methods of use thereof
EP2011498A4 (fr) * 2006-04-10 2009-11-11 Mitsubishi Gas Chemical Co Agent ameliorant les fonctions cerebrales et aliment fonctionnel contenant l'agent ameliorant
US10531655B2 (en) 2011-12-02 2020-01-14 The Regents Of The University Of California Reperfusion protection solution and uses thereof
EP3090738A1 (fr) * 2015-05-04 2016-11-09 Universidade de Santiago de Compostela Riboflavine pour le traitement d'un accident ischémique et/ou d'autres maladies associés à l'excitotoxicité du glutamate
WO2016177840A1 (fr) * 2015-05-04 2016-11-10 Universidade De Santiago De Compostela Riboflavine pour le traitement d'un accident ischémique cérébral et/ou d'autres maladies associées à l'excitotoxicite du glutamate
JP2018131410A (ja) * 2017-02-15 2018-08-23 ヒノキ新薬株式会社 カスパーゼ−３阻害剤とその用途

Also Published As

Publication number Publication date
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