Method and composition for treating mammalian diseases caused by inflammatory response.

one. Field of the Invention

This invention relates to therapeutic methods for preventing and treating damage and the resulting disease state in mammals caused by mammalian cells involved in the inflammatory response resulting in undesirable respiratory explosion, production of enzymes and cellular signaling agents in mammalian cells. This invention also relates to compositions used in therapeutic methods.

Description of the Prior Art

Active oxygen species are generated by cells in response to inter alia aerobic metabolism, drug catabolism, toxins and other cenobites, ultraviolet light and X-ray radiation, and the respiratory explosion of phagocytic cells (such as white blood cells) to kill invasive bacteria and in response to foreign bodies. Hydrogen peroxide, for example, is produced during the respiration of most living organisms, especially by living, depressed cells.

These species of active oxygen can injure cells. An important example of such damage in lipid peroxidation involving the oxidative degradation of unsaturated lipids. Lipid peroxidation is highly damaging to the structure and functions of membranes and can cause numerous cytopathological effects. Cells defend against lipid peroxidation by producing scavenging radicals such as superoxide dismustase, catalase, and peroxidase. Injured cells have a decreased ability to produce radical cleaners. Excess hydrogen peroxide can react with pyrimidines to open in double bond 5, 6-, This reaction inhibits the ability of pyrimidines to bind hydrogen with complementary bases, Hallender et al. (1971). Such oxidative biochemical injury can result in loss of cell membrane integrity, reduced enzyme activity, changes in transport kinetics, changes in lipid content of the membrane, and leakage of potassium ions, amino acids, and other cellular materials.

The production of active oxygen intermediates has been suggested to cause many disorders of the skin, tissue, and organs such as atherosclerosis, arthritis, cytotoxicity, skin inflammation, photoaging, wrinkles, actinic keratosis, tumor formation, cancer, hypertension , Parkinson's disease, lung disease and heart disease. The role of active oxygen radicals in tumor promotion has been proposed based on the findings that 8a) tumor promoters increase the level of oxygen radicals, (b) many free radical generating systems promote tumors, and (c) certain Antioxidants inhibit the biochemical effects of tumor promoters.

In vitro, reactive oxygen intermediates can be generated in cell culture medium by autoxidation and photo-oxidation of medium components. During excision and storage, transplant organs can suffer oxidative damage that results in loss of integrity in the cell membrane and shortens the life of the organ.

When cells are depressed with oxidative damage, a resuscitation step is necessary to recondition the cells. Antioxidants have been shown to inhibit damage associated with active oxygen species. For example, pyruvate and other alpha-keto acids have been reported to react rapidly and stoichiometrically with hydrogen peroxide to protect cells from cytolytic effects, O'Donnell-Tomey et al., J. Exp. Med. 165, pages 500-514. (1987).

United States Patent Number issued to Nath presents a method of arresting or preventing acute kidney failure by administering a non-toxic pyruvate salt to a patient in need of such treatment.

Nath's invention provides a therapeutic method comprising administering an amount of pyruvate salt to a patient experiencing or in danger of acute kidney failure. The pyruvate salt, preferably sodium pyruvate, is preferably dispersed or dissolved in a pharmaceutically acceptable liquid carrier and is administered parenterally in an amount effective to arrest or prevent such acute kidney failure, thus allowing restoration of normal kidney function. . In some cases, pyruvate can be applied by infusion directed to the kidney or into the proximal renal arterial circulation. The method is effective in preventing or countering acute kidney failure due to a wide variety of causes, including, but not limited to, traumatic injury, including burn injury and obstruction; Reperfusion after ischemia, inflammatory glomerulonepritis, and sepis, for example, due to negative bacterial infection.

Martin et al., 1994, US Patent No. 5,296,370, presents therapeutic compositions for preventing and reducing injury to mammalian cells and increasing the rate of resuscitation of injured mammalian cells. In one example, the therapeutic compositions comprise (a) a pyruvate chosen from the group consisting of pyruvic acid, pharmaceutically acceptable salts of pyruvic acid and mixtures thereof, (b) an antioxidant and (c) a mixture of saturated fatty acids and unsaturated where fatty acids are those fatty acids required for the restoration of injured mammalian cells.

U.S. Patent No. 5,256,697, issued to Miller et al., Presents a method of orally administering a therapeutically effective amount of pyruvate percussor to a mammal to improve insulin resistance, lower levels of long-lasting insulin, and reduce fat gain.

US Patent Nos. 3,920,835, 3,984,556, and 3,988,470, all issued to Van Scott et al., Disclose methods of treating acne, dandruff, and palmar keratosis, respectively, which consist of applying to the affected area a topical composition comprising about 1 % to about 20% of a minor aliphatic compound containing from two to six carbon atoms chosen from the group consisting of a20 keto acids, α-keto acids and esters thereof and 3-hydroxybutyric acid in a pharmaceutically acceptable carrier. Aliphatic compounds include pyruvic acid and lactic acid.

US Patent Nos. 4,105,783 and 4,197,316, both issued to Yu et al., Disclose a method and a composition, respectively, for treating dry skin consisting of the application to the affected area of ​​a topical composition comprising from about 1% to about 20% of a compound chosen from the group consisting of amides and ammonium salts of 24 ¿/ iz-hydroxy acids and Be / cz-hydroxy acids and? l // iz-keto acids in a pharmaceutically acceptable carrier. Compounds include amides and ammonium salts of pyruvic acid and lactic acid.

U.S. Patent No. 4,234,599, issued to Van Scott et al., Presents a method of treating actinic and non-actinic epidermal keratoses by applying a topical composition to the affected area that comprises an effective amount of a compound chosen from the group consisting of of J / fa-hydroxy acids, / J-hydroxy acids and Alpha-keto acids in a pharmaceutically acceptable carrier. Acidic compounds include pyruvic acid and lactic acid.

United States Patent Number 4,294,852, issued to Wildnauer et al., Describes a skin treatment composition comprising the JZ / iz-hydroxy acids, β hydroxy acids and / IZ / a-keto acids discovered above by Van Scott et al., In combination with aliphatic alcohols C₃-C₈.

US Patent No. 4,663,166, issued to Veech, discloses an electrolytic solution comprising a mixture of L-lactate and pyruvate in a ratio of 20: 1 to 1: 1, respectively, or a mixture of DP-hydroxybutyrate and acetoacetate, in a ratio of 6: 1 and 0.5: 1, respectively.

Sodium pyruvate has been reported to reduce the number of erosions, ulcers, and bleeding in the gastric mucosa in guinea pigs and rats caused by acetylalicyl acid. The analgesic and antipyretic properties of acetylacylilic acid were not altered by sodium pyruvate, Puschmann, Arceneimittelforschung, 33, pages 410-415 and 415-416 (1983).

Pyruvate has been reported to exert a positive inotropic effect on stunned myocardium, which is prolonged ventricular dysfunction after brief periods of coronary artery occlusion without causing irreversible damage, Mentzer, et al., Ann. Surg., 209, page 629-633 (1989).

Pyruvate has been reported to produce a relative stabilization of left ventricular pressure and work parameter and reduce the size of heart attacks. Pyruvate improves the summary of the spontaneous heart rhythm and the restoration of normal rhythms and pressure development, Bunger et al., J. Mol. Cell. Cardiol., 18, pages 423-428 (1986), Mochizuki et al., J. Physiol. (Paris), 76, page 805-812 (1980), Regitz et al., Cardiovasc. Res., 15, pages 652-657 (1981), Giannelli et al., Ann. Thorac. Surg., 21, pages 386-396 (1976).

Sodium pyruvate has been reported to act as an antagonist of cyanide poisoning (presumably through the formation of a cyanohydrin) and protect against the lethal effects of sodium sulfide and delay the activation and development of functional, morphological measures and biochemistry of axons acrylamide neuropathy,

Schwartz et al., Toxicol, Appl. Pharmacol., 50, pages 437-442 (1979), Sabri et al., Barin Res., 483, pages 1-11 (1989).

A chemotherapeutic cure for advanced L1210 leukemia has been reported 5 using sodium pyruvate to restore abnormally deformed red blood cells to normal. Deformed red blood cells prevent proper drug delivery to tumor cells, Cohen, Chemother Cancer.

Pharmacol., 5, pages 175-179 (1981).

Primary cultures of heterotopic tracheal transplants exposed in vivo to 7,

12-dimethyl-benz (a) anthracene were reported to have been maintained in enriched media supplemented with sodium pyruvate together with cultures of peripheral blood lymphocytes stimulated with interlecin-2 and plasmacytomas and hybridomas, pig embryos and human blasts, Schacter, J Immunol. Methods, 99, pages 259-270 (1987), Marchok et al., Cancer Res., 37, pages 1811-1821 (1977), Davis, J. Repord. Fertile. Suppl., 33, pages 115-124 (1985), Okamoto et al., No To Shinkei, 38, pages 593-598 (1986), Cohén et al., J. In Vitro Fert. Embryo Transfer, 2, pages 59-64 (1984).

US Patents numbers 4,158,057, 4,351,835, 4,415,576 and

4,645,764, all awarded to Stanko, discover methods to prevent the accumulation of fat in the liver of a mammal due to the ingestion of alcohol, to control weight in the mammal, to inhibit body fat while increasing the protein concentration in a mammal and to control the disposition of body fat in a living being, respectively. The methods comprise administering to a mammal a mixture

-8δ therapeutic pyruvate and dihydrooxyacetone and optionally rivoflavin. US Patent No. 4,548,937, issued to Stanko, discloses a method of controlling the weight gain of a mammal which comprises administering to the mammal a therapeutically effective amount of pyruvate and optionally riboflavin. US Patent No. 4,812,479, issued to Stanko, describes a method of controlling weight gain in a mammal comprising administering to the mammal a therapeutically effective amount of dihydroxyacetone and optionally riboflavin and pyruvate.

Rats fed a calcium oxalate lithogenic diet including sodium pyruvate were reported to develop fewer urinary stones (stones) than control rats not fed sodium pyruvate, Ogawa et al., Hinyokika Kiyo, 32, pages 1341-1347 (1986).

US Patent No. 4,521,375, issued to Houlsby, describes a method of sterilizing surfaces that come into contact with living tissue. The method comprises sterilizing the surface with aqueous hydrogen peroxide and then neutralizing the surface with pyruvic acid.

US Patent No. 4,416,982, issued to Tauda et al., Discloses a method of decomposing hydrogen peroxide by reacting hydrogen peroxide with a phenol or aniline derivative in the presence of peroxidase.

US Patent No. 4,696,917, issued to Lindstrom et al., Discloses an ocular irrigation solution comprising Eagle's Minimum Essential Medium with Eagle's salts, chondroitrine sulfate, a buffer solution, 2-mercaptoethanol and a pyruvate. The irrigation solution may contain as an option ascorbic and a5 tocopherol. U.S. Patent No. 4,725,586, issued to Lindstrom et al., Discloses an irrigation solution comprising a balanced saline solution, chondroitin sulfate, a buffer, 2-mercaptoethanol, sodium bicarbonate or dectrose, a pyruvate, a system regulator of sodium phosphate and cystine. The irrigation solution may optionally contain ascorbic acid and gamma-tocopherol.

United States Patent No. 4,847,069, issued to Bissett et al., Discloses a photoprotective composition comprising (a) a sorbohydroxamic acid, (b) an anti-inflammatory agent chosen from steroidal anti-inflammatory agents and a natural anti-inflammatory agent and (c ) a topical carrier. There may be fatty acids present as an emollient. U.S. Patent No. 4,847,071, issued to Bissett et al., Discloses a photoprotective composition comprising (a) a radical ester tocopherol or tocopherol expeller, (b) an anti-inflammatory agent chosen from stereoidal anti-inflammatory agents and an anti-inflammatory agent. natural inflammatory and (c) a topical carrier. US Patent No. 4,847,072, issued to Bissett et al., Discloses a topical composition comprising no more than 25% tocopherol sorbate in a topical carrier.

The addition of sodium pyruvate to bacterial and yeast systems has been reported to inhibit the production of hydrogen hydroxide, increase growth, and protect systems against the toxicity of active oxygen intermediates. Acids

-1010 saturated fats contained within chicken fat improved membrane repair and reduced cytotoxicity. The antioxidants glutathione and thioglycolate reduced the damage induced by radical oxygen species, Martin, Ph, D. thesis, (19871989).

While the above compositions and therapeutic methods have been reported to inhibit the production of reactive oxygen intermediates, none of the compositions and methods treat the resulting damage and disease state in mammals caused by undesirable respiratory bursts, the production of enzymes and agents of cellular signaling in mammalian cells.

SUMMARY PE THE INVENTION

The present invention relates to a method of treating the disease state in mammals caused by mammalian cells involved in the inflammatory response and compositions useful in the method. The method of treating mammalian disease status caused by mammalian cells involved in the inflammatory response comprising: contacting the mammalian cells involved in the inflammatory response with an inflammatory mediator; wherein the inflammatory mediator is present in an amount capable of reducing the undesirable inflammatory response and an antioxidant.

The inflammatory mediator, in addition to reducing the undesirable inflammatory response and being an antioxidant, can also provide a source of cellular energy and be a building block in the cellular synthesis of other cellular components. The inflammatory mediator can also increase cellular metabolic rate.

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The present invention also relates to compositions for reducing and treating the disease state in mammals caused by the undesirable inflammatory response comprising: A mediator of the inflammatory response; and a carrier composition; wherein the mediator of the inflammatory response is an antioxidant and capable of reducing the undesirable inflammatory response in mammalian cells.

The inflammatory response mediators can be used individually, in combination and in addition in combination with a therapeutic agent such as an antibacterial, antiviral, antifungal, protein, enzyme, antihistamines, hormones, non-steroidal anti-inflammatory, cytokine and steroid.

A preferred method of administering the inflammatory mediator is inhalation.

DETAILED DESCRIPTION PE THE INVENTION

Therapeutic compositions and a method of treating the mammalian disease state caused by mammalian cells involved in the inflammatory response have been discovered. The mammalian cells primarily responsible for the inflammatory response are white blood cells or leukocytes.

In a method of treating mammalian disease status caused by mammalian cells involved in the inflammatory response, mammalian cells are brought into

-1212 contact with an inflammatory mediator. The inflammatory mediator is present in an amount capable of reducing the undesirable inflammatory response and is an antioxidant.

The inflammatory response, often referred to as a respiratory burst, is the response of defensive mammalian cells, primarily white blood cells or leukocytes. These cells normally respond to mammalian injury or invasion by releasing a number of active compounds at the site of injury or invasion. Among the released compounds are enzymes like proteases and active oxygen species like hydrogen peroxide.

One purpose of the respiratory blast is to provide a battery of oxidative agents in response to the stimulant that can be used by leukocytes for the destruction of foreign cells, viruses, particles, and some toxins that have been ingested or are in the vicinity of the leukocyte. The term "respiratory blast" refers to a coordinated series of metabolic events that take place when leukocytes are exposed to appropriate stimuli. This group of events supports all oxygen dependent deaths by leukocytes.

The first of these events is the sharp increase in oxygen absorption that occurs after leukocyte stimulation. While the oxygen consumption of resting leukocytes varies widely by cell type, they all respond to appropriate stimuli with increased oxygen absorption.

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Leukocyte stimulation also causes an increase in glucose oxidation through the passage of hexose monophosphate. The hexose monophosphate step is a metabolic pathway in which glucose is oxidized to carbon dioxide and a five carbon sugar, with NADP + serving as an electron acceptor. Activation of the hexose homophosphate passage therefore means that the oxidation of NADPH to NADP + is increased during the respiratory burst.

The respiratory explosion produces superoxide and hydrogen peroxide. The oxygen taken in by the respiratory blast is converted to superoxide. Hydrogen peroxide seems to increase during the respiratory explosion mainly from the stimulation of the superoxide anion.

0-2 + ​​O2 + 2H 'H2 O2 + O2

It has been demonstrated by Root and Metcalf and reported in J. Clin. Invest., 60: 1266 that 15 80 percent of the superoxide is converted to hydrogen peroxide and the disinstimulation reaction is the only major source of the hydrogen peroxide generated during the explosion. Hydrogen peroxide and superoxide are considered responsible for the death of leukocytes.

Many agents, both soluble and particulate, are capable of triggering the respiratory blast. Particulate activating agents include bacteria, viruses, and fungi for internal body organs or areas, and bacteria, viruses, fungi, fibers, smoke, dust, ash, pollen, smog, and the like for body cavities and organs such as the lungs, skin, digestive tracts. and execratory open to the environment. Soluble agents can be

-1414 toxins, medicinal compounds and soluble excretions of infected bacteria, fungi and mammalian cells and the like.

Activation of the respiratory burst in leukocytes normally follows exposure to stimuli for less than a minute. After stimulation of the respiratory burst, the consumption of oxygen in the leukocytes increases by more than 100 times, which results in, among other things, the production of superoxide, peroxide and hydrogen peroxide. The term "leukocytes" as used herein includes lemocytes, phagocytes, macrophages, and helper cells.

Usually, after the respiratory explosion, the stimulant and / or the stimulation mechanism is switched off, allowing the leukocyte to return to its normal resting state. When the explosion does not stop, the inflammatory action of leukocytes continues unchecked causing a number of disease states. These disease states 15 occur as the compounds produced by the attack of leukocytes injure and kill tissue cells and other leukocytes. It is this failure to terminate the respiratory blast and the resulting injury to surrounding tissue cells, blood cells, other leukocytes, and injured cells that causes the disease states treated with the present invention. The undesirable inflammatory response occurs when the inflammatory response causes damage to host cells and this injury poses an independent threat to the host.

In a preferred example, therapeutic compositions containing an inflammatory mediator are administered locally to the site of inflammation. In another preferred specimen, the therapeutic compositions are administered systemically. In

-1515 yet another favorite specimen, the therapeutic compositions are administered systemically and locally together.

In a preferred example, the therapeutic compositions are administered by inhalation. The therapeutic compositions can be nebulized first by any appropriate means. The therapeutic compositions can be in liquid or solid form with liquid pills or particle sizes that are small enough to facilitate access to the lung tissue by inhalation.

In another preferred specimen, a sterile therapeutic agent solution is nebulized and inhaled by the patient. A therapeutically effective amount of medically inflammatory inhalation. This can be accomplished in a single inhalation or by repeated inhalations over a period of time typically 1 to 30 minutes. Preferably inhalation will be complete in less than 20 minutes. Most preferably, the inhalation will be complete in less than 15 minutes.

The term "injured cell" as used herein means a cell that has some or all of the following: (a) damaged membranes so that transport across the membrane is decreased resulting in an increase in toxins and normal cellular waste within the cell and a decrease in nutrients and other components necessary for cellular repair within the cell, ( b) an increase in the concentration of oxygen radicals within the cell due to the decreased ability of the cell to produce antioxidants and enzymes or (c) damaged DNA, RNA and ribosomes that s

-1616 must repair or replace before normal cellular functions can be continued.

Preferably the inflammatory mediator when contacted with a mammalian cell provides a source of cellular energy and a building block in the cellular synthesis of other cellular components.

The inflammatory response that is reduced is at least one of the following: oxygen radical production, peroxide production, cytokine and / or protease production, prostiglandin production, erythema, histamine and interlukin production, and similar responses known in the art as responses inflammatory.

The preferred inflammatory mediator is at least one compound chosen from the group consisting of a pyruvate percussor, pyruvate, a lactate percussor, and lactate. A percussor is a substance from which another substance is formed and in this text includes salts.

Preferably pyruvate is chosen from the group consisting of pyruvic acid, lithium pyruvate, potassium pyruvate, magnesium pyruvate, calcium pyruvate, zinc pyruvate, manganese pyruvate, and the like, and mixtures thereof. Sodium pyruvate is the most preferred.

Other preferred inflammatory mediators are selected from the group consisting of pyruvilglycene, pyruvil-alanine, pyruvil-leucine, pyruvil-valine, pyruvil-isoleucine, pyruvil-phenylalanine, pyruvamide, dihydroxyacetone, and propylene glycol.

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The preferred salts of the inflammatory mediator are salts that do not have an adverse effect on the mammalian cell when applied as a salt of the inflammatory mediator. Typical salts would be lithium, sodium, potassium, aluminum, magnesium, calcium, zinc, manganese, ammonia, and the like, and mixtures thereof.

The lactate percussor is preferably chosen from the group consisting of lactyl-glycene, lactyl-alanine, lactyl-leucine, lactyl-valine, lactyl-isoleucine, lactyl-phenylalanine, lactamide and the various lactate salts.

Compositions for reducing and treating disease state in mammals caused by undesirable inflammatory response include:

a mediator of the inflammatory response; and a carrier composition.

The carrier composition is chosen from the group consisting of tablets, capsules, liquids, isotonic liquids, isotonic medium, enteric, parenteral, topical tablets, capsules, creams, gels, ointments, chewing gums, confectionery and the like.

The inflammatory mediator is administered in a therapeutically effective amount to reduce the undesirable inflammatory response. Preferably from 0.001 grams to 10 grams per dose. More preferably 0.001 to 1 gram per dose and more preferably 0.001 to 0.25 grams per dose. It is understood that the method of administration and the condition to be treated will greatly affect the dose required to achieve the therapeutic effect.

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Typical airway diseases treatable with the present compositions and method include, but are not limited to, bronchial asthma, acute bronchitis, emphysema, chronic obstructive emphysema, centrilobular emphysema, pancinar emphysema, critical obstructive bronchitis, reactive air disease, cystic fibrosis, bronchistasis, acquired bronchiestais, kartaagener syndrome, atelectasis, acute atelectasis, chronic atelectasis, pneumonia, essential thrombocytopenia, Legionnaires' disease, psittacosis, fibrogenic dust disease, disease due to organic dust, diseases due to irritating gases and chemicals, hypersensitivity diseases of the lung, idiophatic infiltrative diseases of the lungs and the like.

The disease states to treat in particular are emphysema and asthma.

The inflammatory mediator of the present invention can be administered before, after, and / or with other therapeutic agents. Typical therapeutic agents are antibacterial, antiviral, antifungal, antihistamines, proteins, enzymes, hormones, non-steroidal anti-inflammatory drugs, cytokines, steroids, and the like.

Obviously, numerous modifications and variations of the present invention are possible in light of the above teachings and the invention is not limited to the example herein. It is therefore understood that within the scope of the appended claims, the invention may be practiced in a manner other than that specifically described herein.

EXAMPLE

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Subject: A 59-year-old man suffering from emphysema and restrictive air disease was treated as described below for three (3) months. Prior to treatment, the subject had limited ability to breathe, was unresponsive to any other treatment, required daily oxygen and was unable to function at work (had sabbatical leave). After three (3) months of treatment showed marked improvements. In fact, dramatic results were observed within two (2) weeks.

Treatment: Treatment was as follows:

Five (5) milliliters of five (5) millimolar sodium pyruvate solution is sterilized by filtration through a 0.1 micron filter. The sterile pyruvate solution is placed in a "Pulmo Aid" nebulizer manufactured by DeVilbiss Co., Somerset, Pennsylvania 15501-0635. The sterile pyruvate solution is nebulized in the Pulmo Aid apparatus, attached to a disposable nebulizer, and inhaled by the patient. The patient inhales normally from the Pulmo Aid nebulizer until all of the solution has been nebulized and inhaled. This inhalation step normally takes approximately ten (10) to twenty (20) minutes.

The patient is treated with this inhalation therapy periodically. Initially, treatments are around four (4) minutes a day with intervals of around six (6) hours. Treatment is reduced to three (3) times a day with intervals of eight (8 = hours after 30 days of therapy. Treatment is further reduced to once a day 60 DAYS AFTER STARTING TREATMENT. After 90 ( 90) days the treatments are three to five times a week.

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The data below shows the results of various lung capacity and lung function tests administered before treatment and two (2) months after treatment started.

<td>Type of test</td><td>Before Treatment</td><td>After two months of treatment</td>

<td>SVC (slow vital capacity)</td><td> 63</td><td> 83</td>

<td>IC (inspiration capacity)</td><td> 64</td><td> 69</td>

<td>ERV (expiration reserve volume)</td><td> 62</td><td> 110</td>

<td>MW (maximum ventilatory volume)</td><td> 19</td><td> 25</td>

<td>DCO (diffusion of carbon monoxide MI / min / mm Hg)</td><td> 27</td><td> 39</td>

<td>DSB (single breath diffusion)</td><td> 42</td><td> 52</td>

<td>O SAT (oxygen saturation)</td><td> 91</td><td> 95</td>

Observation: After twenty or thirty minutes from the start of the treatment, the tidal volume increases, the wheezing stops and an increase in exercise tolerance is observed. Dependence on "Proventil" (Albuterol made by Schering) is reduced from 1600 mg per day to 400 mg per day within the first two (2) weeks of treatment. Oxygen use was immediately discontinued when treatment was started.

Conclusion: The treatment did the following.

(1) Improves lung function by 20%.

(2) Decreased some levels of medications and stopped using oxygen.

(3) Reactive air sickness was reduced to the point that routine use of inhalers is not needed.

-2121 (4) Greater exercise tolerance has been tested and certified as close to that of a person without emphysema and restrictive air disease. The subject has been able to return to full-time work in the previous job.

(5) No recurrence of decreased lung functions.

(6) The mental attitude has greatly improved.

While the method of treating disease status in mammalian cells 10 involved in the inflammatory response described herein constitutes preferred examples of this invention, it should be understood that the invention is not limited to this precise form of method and what changes can be made. therein without departing from the scope of the invention defined in the appended claims.

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