Title:  Herbal remedies for treating allergies and asthma

United States Patent:  6,630,176

Issued:  October 7, 2003

Inventors:  Li; Xiu-Min (Mamaroneck, NY); Sampson; Hugh A. (Larchmont, NY)

Assignee:  Mount Sinai School of Medicine of New York University (New York, NY)

Appl. No.:  800815

Filed:  March 7, 2001

Abstract

The present invention provides herbal compositions that can prevent or reduce the severity, intensity, or duration of allergic and/or asthmatic symptoms and/or can prevent or delay the development of an allergic or asthmatic response to an antigen. The compositions may optionally include one or more adjuvants, cytokines, encapsulating materials, or pharmaceutical carriers or excipients, and may be administered prior to, during, or after the development of allergic or asthmatic symptoms in sensitized individuals. Alternatively or additionally, the compositions may be administered prior to sensitization to a particular antigen; preferably substantially concurrently with exposure to the antigen.

SUMMARY OF THE INVENTION

The invention provides improved treatments for allergies and asthma. In particular, the invention provides herbal formulation compositions that, when administered to an individual suffering from asthmatic or allergic symptoms, reduce the severity, intensity, and/or duration of at least some of those symptoms. Inventive compositions preferably reverse established allergic or asthmatic reactions to particular antigens. Alternatively or additionally, inventive compositions may prevent or delay an allergic or asthmatic reaction and/or may block or reduce the development of allergic or asthmatic sensitivity to antigens.

In one preferred embodiment, an inventive composition comprises one or more components of Fructus Pruni Mume (Wu Mei), Pericarpium Zanthoxyli Bungeanum (Chuan Jiao), Herba cum Radice Asari (Xi Xin), Rhizoma Coptidis (Huang Lian), Cortex Phellodendri (Huang bai), Rhizoma Zingiberis Officinalis (Gan Jiang), Radix Lateralis Aconiti Carmichaeli Praeparata (Fu Zi), Ramulus Cinnamomi Cassiae (Gui Zhi), Radix Ginseng (Ren Shen), and Radix Angelicae Sinensis (Dong gui). Other preferred compositions additionally comprise one or more components of Ganoderma lucidum (Ling Zhi). In a particularly preferred embodiment, the composition comprises the Chinese herbal remedy Wu Mei Wan (WMW) plus Ganoderma lucidum (Ling Zhi, LZ).

In another preferred embodiment, the composition comprises one or more components of Perillae frutescens (su zi), Descurainia Sophia (ting li zi), Raphanus sativus L. (lai fu zi), Marus alba L. (sang bai pi), Prunus armeniaca (xing ren), Scutellaria baicalensis (huang qin), Glycyrrhiza uralensis (gan cao), Ziziphus jujuba (da zao), Aster tataricus (zi wan), Pteria margaritaferae (zhen zhu mu), and Aussilago farfara (kuan dong hua).

In yet another preferred embodiment, the composition comprises one or more components of Perillae frutescens (su zi), Descurainia Sophia (ting li zi), Prunus armeniaca (xing ren), Scutellaria baicalensis (huang qing), Sophora flavescens (ku sen), Angesica sinensis (don gui), Paeonia lactiflora (bai shao), Peuraria lobata (ge gen), Platycodon grandiflorum (jie gen), Pteria margaritaferae (zhen zhu mu), Ganoderma lucidum (ling zhi), Glycyrrhiza uralensis (gan cao), Ziziphus jujuba (da zao), and Frash zingiber officinal (sheng jiang).

In certain preferred embodiments of the invention, inventive compositions are administered in combination with one or more standard therapies. For example, inventive compositions may be administered in combination with corticosteroids (e.g., inhaled, injected, or orally delivered corticosteroids), anti-histamines, decongestants, cromolyn sodium, standard immunotherapy, rush immunotherapy, etc. used to treat allergic or asthmatic symptoms.

Inventive compositions may optionally be characterized in one or more animal model systems.

Furthermore, the present invention provides a method of formulating an herbal remedy of the present invention, and/or of identifying active ingredients in inventive herbal compositions. For example, one or more active components of the herbs of the inventive compositions may optionally be extracted or purified using any technologies known in the art including, but not limited to chromatography, aqueous extraction, organic solvent extraction, etc. The active component and/or whole herbs may be combined with other pharmaceutically acceptable excipients or carriers to make pharmaceutical compositions.

The present invention also provides methods of treating allergies or asthma and/or preventing the development of allergies or asthma. The compositions of the present invention may be administered to an individual to prevent the development of, or reduce symptoms of, an allergic or asthmatic response to an allergen. For example, the inventive compositions may be administered substantially simultaneously with a known allergen in order to alter the immune response so that an allergic or asthmatic response does not develop, or develops to a lesser extent than would be observed in the absence of the inventive composition. In another preferred embodiment, the inventive composition is administered before exposure to a known allergen in order to lessen the allergic or asthmatic reaction. In yet another embodiment, the inventive composition is administered after exposure to a known allergen or after development of sensitization to the allergen in order to lessen the effect of potential allergic or asthmatic reactions in the future.

The present invention also provides methods of identifying and characterizing herbal remedies useful in the treatment of allergic reactions and asthma. In a preferred embodiment, an inventive test herbal remedy is administered to an animal model known to have an immune response similar to that of the individual to be treated with the herbal remedy. The herbal remedy may be administered at any time including, but not limited to, prior to sensitization, during sensitization, after sensitization, prior to challenge, during challenge, or after challenge. In a particularly preferred embodiment, the animal model used is the allergic mouse model. In vitro assays may also be used in characterizing herbal remedies. In a particularly preferred embodiment, a basophil histamine release assay is used in assessing herbal remedies.

In another aspect, the present invention provides methods of identifying herbs useful in preparing herbal remedies and methods of identifying active components of these herbs. Herbs or mixture of herbs are administered to animal models of allergic disease or are used in in vitro assays known to predict effects in vivo. In a preferred embodiment, the active components of the herbs or mixture of herbs are purified or partially purified. Various methods of purification and preparation may be used in providing the formulations to be studied. The prepared/purified formulations may be also used in the various in vitro and in vivo assays described in the present patent application to identify those herbs and active components responsible for treating or preventing allergic disease.

DETAILED DESCRIPTION OF CERTAIN PREFERRED EMBODIMENTS OF THE INVENTION

The present invention unites insights from traditional Chinese medicine and modern Western medicine to formulate treatments for allergy and asthma. Traditional Chinese medicine employs herbal formulations to treat bodily ailments. In some cases, single herbs or herb derivatives are used. More commonly, however, "formulas", or specific combinations of several particular herbs, are administered. The recipes for these formulas are assembled into books known as "formularies". The original formulary, Discussion of Cold-Induced Disorders and Miscellaneous Diseases (Shang Han Za Bing Lun), was written at the end of the second century A.D. by Zhang Zhong-Jing. This book was later edited by Wang Shu-He, who divided it into two parts, Discussion of Cold-Induced Disorders (Shang Han Lun), which deals with externally-contracted diseases, and Essentials from the Golden Cabinet (Jin Gui Yao Lue), which is primarily concerned with internally-generated disorders (Bensky et al., Chinese Herbal Medicine: Formulas & Strategies. Eastland Press, 1999; incorporated herein by reference). These two books contain 374 formulas. The present invention provides a new analysis of certain of these formulas and identifies characteristics applicable to the treatment of asthma and allergic disease according to Western principles.

Several different Chinese formulas are recommended for the treatment of coughing, wheezing, shortness of breath, or other symptoms that can be associated with asthma or allergies. Few, however, have been shown in clinical or animal studies to have a demonstrable salutary effect on asthma or allergies (see, for example, Coyle et al., Eur. J. Pharmacol. 148:51, 1988; But et al., Clin. Rev. Allergy Immunol. 14:253, 1996; Hsieh et al., Pediatr. Allergy Immunol. 14:253, 1996; Zhang et al., Chung. Kuo. Chung. Hsi. I. Chieh Ho. Tsa. Chih. 17:204, 1997; Sun et al., Chung. Kuo. Chung. Hsi. I. Chieh. Ho. Tsa. Chih. 17:201, 1997; Zou et al. Chung Kuo. Chung. Hsi. I. Chieh. Ho. Tsa. Chih. 16:529, 1996; Xu et al., Chung Kuo. Chung. Hsi. I. Chieh. Ho. Tsa. Chih. 16:198, 1996; Chen et al., Pharmacology and Clinical Applications of Patent Medicine and Famous Formulas. Hong Kong: Ya Yi Publishing Company, pp. 44-92, 407, 1989; Egashira et al., Ann. NY Acad. Sci. 685:580, 1993; Noma et al., Arerugi 45:494, 1996; Toda et al., J. Ethnopharmacol. 24:303, 1988; Hamasaki et al., J. Ethnopharmacol. 56:123, 1997; Roberts et al., J. Allergy Clin. Immunol. 82:236, 1988; Dong et al., Chinese J. of Integrated Traditional Chinese and Western Medicine 318, 1989; Nyunt et al., Arerugi 44:503, 1995; each of which is incorporated herein by reference). The present invention identifies herbal formulations, and components thereof, that are useful in alleviating, treating, and/or preventing symptoms of allergy and/or asthma.

Herbal Formulations

The present invention provides herbal formulations that reduce allergic or asthmatic symptoms and signs including but not limited to airway hyperresponsiveness, hives, rash, puritis, watery eyes, bronchconstriction, edema, diarrhea, difficulty breathing, vasodilation, decrease in blood pressure, increased IgE levels, increased plasma histamine levels, increased numbers of goblet cells, increased Th2 cytokine levels, bronchial inflammation, anaphylaxis, and death. Preferably, the compositions demonstrate a reduction in symptoms that is at least as significant as that observed with standard treatments such as corticosteroids and anti-histamines. Preferably, the reduction in symptoms occurs more quickly than is seen with standard treatments (e.g., conventional antigen immunotherapy and/or steroid treatment), is more persistent than that observed with standard treatments, and/or is more extensive than that achieved by standard treatments.

Without wishing to be bound by any particular theory, we propose that inventive herbal compositions down regulate Th2 responses, thereby leading to a reduction in allergy or asthma symptoms. In fact, particularly preferred compositions have specific effects on Th2 responses, rather than general immunosuppressive activities. For example, preferred inventive compositions when compared to immunosuppressive agents such as corticosteroids, FK506, methotrexate, and cyclosporin are more selective for the allergic or asthmatic response.

The current understanding of allergic and/or asthmatic diseases may be used to choose herbal formulations that may be further tested using standard experimental systems used in studying allergies or asthma (for example, see Examples below). Formulations may be chosen which are known to reduce sneezing, water eyes, itching, diarrhea, wheezing, bronchoconstriction, hives, etc, as such symptoms can be associated with allergies and/or asthma The chosen formulation may be assessed in animal models of allergy or asthma, or in in vitro models of allergy or asthma. From the hundreds of herbals formulas known in Eastern medicine, the present invention teaches methods of choosing potential anti-allergy or anti-asthma therapies and testing the chosen therapies for efficacy in treating or preventing allergies or asthma. As described in the Examples below, the present invention also demonstrates use of these methods, and defines certain particularly useful compositions.

For example, according to the present invention, one preferred formulation is Wu Mei Wan, a Chinese medicinal formula. Wu Mei Wan has been used in the treatment of parasitic diseases in Chinese medicine and has also been used to treat endometriosis. Particularly preferred inventive herbal compositions include at least the herbs Fructus Pruni Mume (Wu Mei), Pericarpium Zanthoxyli Bungeanum (Chuan Jia), Herba cum Radice Asari (Xi Xin), Rhizoma Coptidis (Huang Lian), Cortex Phellodendri (Huang Bai), Rhizoma Zingiberis Officinalis (Gan Jiang), Radix Lateralis Aconiti Carmichaeli Praeparata (Fu Zi), Ramulus Cinnamomi Cassiae (Gui Zhi), Radix Ginseng (Ren Shen), and Radix Angelicae Sinensis (Dang gui). If the weight of Pericarpium Zanthoxyli Bungeanum is defined as approximately 1, the ratio of Pericarpium Zanthoxyli Bungeanum to Fructus Pruni Mume is from approximately 10 to approximately 16, to Herba cum Radice Asari is approximately 1, to Rhizoma Coptidis is from approximately 4 to approximately 6, to Cortex Phellodendri is from approximately 3 to approximately 4, to Rhizomo Zingiberis Officinalis is from approximately 3 to approximately 4, to Radix Lateralis Aconiti Carmichaeli Praeparata is approximately 2, to Ramulus cinnamomi Cassiae is approximately 2, to Radix ginseng is from approximately 3 to approximately 4, and to Radix Angleicae Sinensis is from approximately 2 to approximately 3. In certain particularly preferred embodiments, the inventive herbal compositions also include the herb Ganoderma lucidum (ling Zhi). The ratio of the weight of Pericarpium Zanthoxyli Bungeanum to the weight of Ganoderma lucidum is preferably approximately 2. In other preferred embodiments, Radix Codonopsis Pilosulae is substituted for Radix Ginseng. One particularly preferred inventive herbal formulation is the particular formulation of WMW, as described in Table 1. Other preferred compositions are the formulation of WMW and ZN, as described in Table 2.

Other particularly preferred inventive herbal compositions include at least the herbs Huang Qing, Ku Sen, and Ling Zhi. In certain preferred embodiments, these herbs are present in relative amounts approximating 3:3:2 ratios. Alternatively or additionally, preferred compositions may include one or more of Dong Gui and Ge Gen. Relative amounts of these herbs, in certain particularly preferred compositions, will approximate the levels of Huang Qing and/or Ku Sen. Other preferred embodiments may further, or alternatively, include Su Zi, preferably at a level approximating that of Huang Qing and/or Ku Sen. One particularly preferred inventive herbal formulation is MSSM-002 (see Table 3).

Another particularly preferred composition is MSSM-0001 (see Table 4).

Inventive herbal formulations may be prepared in any manner that preserves the desired biological activity of the formulation. Examples of possible preparations include decoctions, aqueous extracts, organic solvent (e.g., alcohol, ethyl acetate) extracts, and dry powder. In one particularly preferred embodiment, the herbs are dried and ground, and the resulting powder is processed into pill form.

Herbs for use in the inventive compositions will generally be provided in their natural, herbal form. The herbs may be harvested from any location at any time of the year. Preferably, the herb has the active components at concentrations sufficient to treat allergic symptoms, treat asthmatic symptoms, or prevent an allergic response. More preferably, the herbs are harvested in a manner which maximizes the efficacy of the herbal formulation. In a particularly preferred embodiment, an herb may be harvested from a specific geographic location or during a particular time of the year to maximize the amount of active ingredient found in the herb. To give but one example, the herb Ganoderma lucidum may preferably be harvested in the Dooryoon Mountains located in the South Cholla Province of Korea.

To one of skill in this art, it will be appreciated that the substrate on which the herbs are growing can be important in selecting the herbs for harvest. For example, the fungi Ganoderma lucidum growing on oak wood may be found to have more of the active ingredient than Ganoderma lucidum growing on other woods. As would be appreciated by one of skill in this art, the soil in which the herbs are growing may also contribute to the concentration of the active ingredient in the harvested herb. In other preferred embodiments, the herbs may be artificially cultivated. For an example of cultivating Ganoderma lucidum, please see U.S. Pat. No. 4,472,907, issued Sep. 25, 1984, incorporated herein by reference.

The herbs for the formulations may also be selected based on any number of criteria including, but not limited to, appearance (e.g., color, texture, etc.), smell, feel, HPLC "finger printing", chromatographic (e.g., HPLC, TLC,GC) fingerprint profiles, presence of a "marker" constituent, etc. In a particularly preferred embodiment, the herbal formulation is prepared by following the FDA's "Guidance for Industry Botanical Products". The herbs may also be checked for the presence of pesticide residues, heavy metal content, etc. to ensure the safety of the final product.

As is appreciated by those skilled in this art, a variety of techniques are well known in the art for extracting, isolating, and/or purifying individual active components of the particular herbs. The present invention encompasses both the identification of such active components as described herein and the incorporation of such components into inventive compositions as described herein.

Purification of Active Components

Individual active components of the herbs or herbal formulations may be identified as described herein and may be isolated and/or purified using any techniques known in the art. The active component may be purified from the herb itself in any form (e.g., fruit, seed, spore, flower, leaves, stalk, root, rhizomes, etc.), the culture media of the organism, the decoction of WMW, the decoction of WMW+LZ, etc. Various techniques that may be employed in the purification include filtration, selective precipitation, extraction with organic solvents, extraction with aqueous solvents, column chromatography, high performance liquid chromatography (HPLC), etc. (Zubrick, The Organic Chem Lab Survival Manual Third Edition New York: John Wiley & Sons, Inc., 1992; Scopes Protein Purification Principles and Practice (2nd ed.), New York: Springer-Verlag, 1987; each of which is incorporated herein by reference). As would be appreciated by one of skill in the art, the active components may be proteins, peptides, nucleic acids, natural products, terpenes, alkaloids, proteoglycans, polysaccharides, lipids, triglycerides, etc., and therefore, the purification procedure would depend on the nature of the component being purified.

Fractions from a purification step may be assayed for the desired biological activity to determine which of the fractions contain the desired active component. In a preferred embodiment, the allergic mouse model as described in U.S. patent application Ser. No. 09/518,346, filed Mar. 3, 2000, incorporated herein by reference, is used as the assay for determining the presence of the desired active component. In animal models, administration of the herb or active component may lead to at least a 10% decrease in IgE levels, more preferably at least a 50% decrease, and most preferably at least a 75% decrease. In another particularly preferred embodiment, antigen-specific IgE levels are less than 2000 ng/ml after administration of the inventive formulation, more preferably less than 1500 ng/ml, and most preferably less than 1000 ng/ml. In other preferred embodiments, cell culture based assays or in vitro assays are used (e.g., basophil histamine release assays. For example, in the basophil histamine release assay the herb or active component may lead to at least a 25% decrease in histamine levels; more preferably at least a 50% decrease; and most preferably at least a 75% decrease. As appreciated by one of skill in this art, the desired biological activity may not lie in one fraction by itself but may stem from a combination of active components so various combinations of fractions from each purification step may need to be evaluated in the assay system to identify the active components.

In yet other preferred embodiments, the presence and purity of the active compound is assessed by chemical methods including nuclear magnetic spectroscopy (NMR), mass spectroscopy, infrared spectroscopy (IR), ultra-violet visible spectroscopy, elemental analysis, polarimetry, refractometry, etc.

In the final composition to be delivered to the individual, the purified active component is preferably greater than 50% pure. In a preferred embodiment, the active component is greater than 75% pure, and more preferably greater than 90% pure. In a particularly preferred embodiment, the active component is greater than 95% pure.

Other Components

As will be appreciated by those of ordinary skill in the art, the inventive herbal formulations may be desirably combined with one or more additional components, in a single composition or in more than one composition, to more effectively treat allergic or asthmatic conditions. For example, inventive herbal compositions may be combined with one or more adjuvants, cytokines, or encapsulating materials as discussed more fully below. Additionally, inventive herbal compositions may be combined with other known allergy or asthma treatments such as, for example, general immunosuppressants (e.g., corticosteroids), anti-histamines, cromolyn sodium, traditional immunotherapy, rush immunotherapy, etc.

ADJUVANTS

A variety of compounds are known in the art to have specific or general immunostimulatory effects. Such compositions are commonly referred to as "adjuvants". A large number of adjuvant compounds is known; a useful compendium of many such compounds is prepared by the National Institutes of Health and can be found on the world wide web (http:/www.niavd.nih.gov/daids/vaccine/pdt/compendium/pdf, incorporated herein by reference; see also Allison Dev. Biol. Stand. 92:3-11, 1998; Unkeless et al. Annu. Rev. Immunol. 6:251-281, 1998; Phillips et al. Vaccine 10:151-158,1992; each of which is incorporated herein by reference). Preferred adjuvants are characterized by an ability to stimulate Th1 responses preferentially over Th2 responses and/or to down-regulate Th2 responses. In fact, in certain preferred embodiments of the invention, adjuvants that are known to stimulate Th2 responses are avoided. Particularly preferred adjuvants include, for example, preparations (including heat-killed samples, extracts, partially purified isolates, or any other preparation of a microorganism or microorganism component sufficient to display adjuvant activity) of microorganisms such as Listeria monocytogenes or others (e.g., Bacille Calmette-Guerin [BCG], Corynebacterium species, Mycobacterium species, Rhodococcus species, Eubacteria species, Bortadella species, and Nocardia species), and preparations of nucleic acids that include unmethylated CpG motifs (see, for example, U.S. Pat. No. 5,830,877; and published PCT applications WO 96/02555, WO 98/18810, WO 98/16247, and WO 98/40100, each of which is incorporated herein by reference). Other preferred adjuvants reported to induce Th1-type responses and not Th2-type responses include, for example, Aviridine (N,N-dioctadecyl-N'N'-bis (2-hydroxyethyl)propanediamine) and CRL 1005.

In some embodiments of the invention, the adjuvant is associated (covalently or non-covalently, directly or indirectly) with the herbal formulation so that adjuvant and formulation can be delivered substantially simultaneously to an individual, optionally in the context of a single composition. In other embodiments, the adjuvant is provided separately. Separate adjuvant may be administered prior to, simultaneously with, or subsequent to herbal formulation administration. In certain preferred embodiments of the invention, a separate adjuvant composition is provided that can be utilized with multiple different herbal formulations.

Where adjuvant and formulation are provided together, any association sufficient to achieve the desired immunomodulatory effects may be employed.

CYTOKINES AND INDUCING AGENTS

In some cases, in will be desirable to provide inventive herbal formulations in combination with one or more cytokines or inducing agents, preferably to promote and/or reflect a reduction in Th2 responses and/or an increase in Th1 responses to the relevant antigen. In certain preferred embodiments of the invention, herbal formulations are provided in combination with one or more Th1 stimulating cytokines (e.g., IL-12, IL-2, IL-18, IL-1.beta. or fragments thereof, IFN.alpha., and/or IFN.gamma., etc.) and/or one or more Th1 inducing agents (e.g., factors such as LPS, CD40, CD40 ligand, BCGs, oligonucleotides containing CpG motifs, TNF.alpha., and microbial extracts such as preparations of Staphylococcus aureus, heat killed Listeria, etc.). Alternatively or additionally, the herbal formulations may be provided in combination with one or more Th1 cytokines (e.g., IL-10, IL-2, IL-12, IL-18, IFN.alpha., IFN.gamma., TNF.beta., etc.).

STANDARD THERAPIES

Inventive herbal formulations may be administered to a subject in combination with one or more other therapeutic treatments. For example, corticosteroid administration is an established and accepted treatment for asthma; inventive herbal formulations may desirably be administered in combination with standard or reduced corticosteroid treatments, whether inhaled or systemic. In the case of treating individuals with allergies, the inventive composition may be administered in combination with accepted treatments for allergies including, but not limited to, anti-histamines, non-steroidal anti-inflammatory drugs, decongestants, and cromolyn sodium. The inventive herbal formulations may also be administered in combination with standard immunotherapy or rush immunotherapy. Immunotherapies are typically administered in order to induce tolerance in a sensitized individual (for a more detailed description of immuotherapy, please see U.S. Provisional Patent Application, U.S. Ser. No. 60/213,765, filed Jun. 23, 2000; incorporated herein by reference).

ENCAPSULATION

Inventive herbal formulations may be administered, whether alone or in combination with one or more other agents or compounds, in the context of an encapsulated system. A variety of encapsulation systems are known in the art (see, for example, discussions in U.S. Ser. No. 60/169,330, filed Dec. 6, 1999, and incorporated herein by reference); any such system may be employed in accordance with the present invention. In certain preferred embodiments of the invention, the encapsulation material itself may offer adjuvant activity. Also, preferred encapsulation systems may desirably be associated with one or more targeting agents that facilitate delivery of the inventive compositions to relevant sites (e.g., mucosal membranes).

PHARMACEUTICAL EXCIPIENTS AND CARRIERS

Pharmaceutical compositions for use in accordance with the present invention may include a pharmaceutically acceptable excipient or carrier. As used herein, the term "pharmaceutically acceptable carrier" means a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Some examples of materials which can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as propylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

Preferably, the inventive pharmaceutical compositions comprising herbal formulations are administered orally. However, other routes of administration may also be utilized. For example, in some embodiments of the invention, pharmaceutical compositions may be delivered to mucous membranes, for example, by inhalation or injection. In general, inventive pharmaceutical compositions can be administered to humans and/or to other animals, orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), bucally, or as an oral or nasal spray.

Preferred oral forms for administration of inventive herbal formulations are described in standard herbal remedies texts (see, for example, Bensky et al., Chinese Herbal Medicine: Formulas & Strategies Eastland Press, 1999; incorporated herein by reference). However, other forms may alternatively be useful. Techniques for preparing alternative forms of pharmaceutical compositions are well known in the art. Several non-limiting examples are discussed, for example, in U.S. patent application Ser. No. 09/518,346, entitled "Animal Model for Allergies", filed Mar. 3, 2000, and incorporated herein by reference.

Identification and Characterization of Inventive Herbal Formulations

The effects of inventive herbal formulation compositions may be studied in humans or in any available in vivo or in vitro model system. Animal models are particularly useful for the identification, characterization, and analysis of a particular composition's effects. Ideally, a model system should reflect closely at least some aspect of the disease pathology in man (or in another organism to which an inventive composition is to be administered for the treatment of asthma or allergy), should be reliable and reproducible, should allow objective measurements of one or more physiologically-relevant parameters, should respond to one or more known therapeutic agents in a manner similar to that observed in man (or the suffering organism), and/or should offer a large number of reagents with which the immune system can be analyzed.

A variety of animal models, including those in guinea pigs, rabbits, sheep, dogs, monkeys, and mice have been developed that can usefully be employed to characterize herbal compositions of the present invention (see, for example, Kay (ed.) Allergy and Allergic Diseases Blackwell Science, Ltd., Oxford. pp. 1037-1110, 1997; Ermel et al. "The Atopic Dog: A Model for Food Allergy" Lab. Animal Science 47(1):40-49, 1997; Li et al. "A Murine Model of IgE-Mediated Cow's Milk Hypersensitivity" J. Allergy Clin. Immunol. 103(2 Pt 1):206-214, 1999; Li et al. "Strain-Dependent Induction of Allergic Sensitization Caused by Peanut Allergen DNA Immunization in Mice" J. Immunol. 162(5):3045-3054, 1999; McCaskill et al. "Anaphylaxis Following Intranasal Challenge of Mice Sensitized with Ovalbumin" Immunology 51:669-677, 1984; U.S. patent application Ser. No. 09/518,246, filed Mar. 3, 2000; each of which is incorporated herein by reference).

To give but one example, guinea pigs were one of the earliest asthmatic model systems because of the ease with which they can be sensitized to foreign antigens, and the similarity of the histological characteristics observed in antigen-sensitized guinea pig lungs as compared with asthmatic humans lungs (Kallos et al., Int. Arch. Allergy Appl. Immunol. 73:77, 1985; incorporated herein by reference). Antigen challenges to sensitized guinea pigs can provoke both early and late-phase airway responses, and the roles of IL-5 and eotaxin in asthmatic reactions have been extensively characterized in this model (Rothenberg et al., J. Exp. Med. 181:1211, 1995; incorporated herein by reference). However, the guinea pig may not be an ideal model since several compounds that have shown therapeutic efficacy in the guinea pig have proven not to be useful in humans, and vice versa (see, for example, Mishall et al., in Allergy and Allergic Diseases (Kay, Ed.) Blackwell Science, Ltd., Oxford, pp. 1037-1110, 1997; incorporated herein by reference).

Mouse models are particularly preferred for use in the characterization of inventive herbal compositions for the treatment of asthma and allergy. The immune system of the mouse mimics the human immune system more closely than does that of other rodents. Furthermore, the mouse immune system has been well characterized through the close analysis of highly inbred strains. In addition, a wide variety of immunological reagents have been developed for use in the analysis of murine immunological reactions, and increasing numbers of useful knock-out and transgenic strains (including, for example, IL-4 deficient mice [Kopf et al., Nature 362:245, 1993, incorporated herein by reference], IgE-deficient mice [Oettgen et al., Nature 7370:367, 1994, incorporated herein by reference], etc.) have been created.

Those of ordinary skill in the art will recognize that the particular mouse strain or route of administration of sensitizing antigen may not be critical in developing a preferred mouse model system for use in characterizing inventive herbal compositions. For example, Renz et al. have described a BALB/c mouse sensitized with aerosolized ovalbumin over a 10-day period (Renz et al., J. Exp. Med. 177:1175, 1993; incorporated herein by reference). These mice show elevated levels of ovalbumin-specific IgE and infiltration of eosinophils into the airway following bronchial challenge. Wills-Karp et al. have described an asthmatic A/J mouse model sensitized by intraperitoneal administration of antigen, followed by intratracheal challenge (Gavett et al., Am. J. Respir. Cell Mol. Biol. 10:587, 1994; Keane-Myers et al., J. Immunol. 161:919, 1998; Wills-Karp et al., Science 282:2258, 1998; Grunig et al., Science 282:2261, 1998; each of which is incorporated herein by reference). Preferably, the sensitizing antigen is administered to the animal via the same route the animal would encounter the allergen in nature (e.g., oral for food allergens, IV or parenteral for venoms, inhaled for pollens or dust allergens, intradermal for latex). In a particularly preferred embodiment, the mouse is sensitized to the allergen using alum as an adjuvant (see Example 1 below for details).

We herein describe (see Example 1) a conalbumin-allergic AKR/J mouse in which IgE significantly increased following intraperitoneal sensitization, and BALF eosinophils were increased at 12 hour and peaked at 72 hours following i.t. challenge. This model closely mimics the late phase response of human asthma and is particularly preferred for use in the characterization of inventive herbal compositions (Li et al., J. Immunol. 160:1378, 1998; incorporated herein by reference). Those of ordinary skill in the art will appreciate that any of a variety of other mouse systems can be developed and/or utilized in accordance with the present invention.

A mouse model using alum as the adjuvant in sensitizing the mouse may also be used in characterizing the herbal compositions of the present invention. The alum mouse is a well-characterized animal model used in studying allergies (Levine et al. "Effect of combinations of inbred strain, antigen, and antigen dose on immune responsiveness and reagin production in the mouse. A potential mouse model for immune aspects of human atopic allergy" Int. Arch. Allergy Appl. Immunol. 39(2-3):156-171, 1970; incorporated herein by reference).

One in vitro model useful in characterizing inventive herbal formulations is the basophil histamine release assay. One of the way in which inventive herbal formulations may be characterized is by their ability to inhibit histamine release in isolated basophils that are contacted with antigen. Example 5 describes one procedure by which such basophil histamine release is assayed; those of ordinary skill in the art will recognize that various modifications and alterations of this precise procedure can be made without departing from the spirit or scope of the present invention. Basophil histamine release assays are well established in the art (to give but a few examples, see Counsell et al., J. Allergy Clin. Immunol. 98:884, 1996; Haselden et al., J. Exp. Med. 189:1885, 1999; incorporated herein by reference).

Uses

The inventive compositions may be employed to treat existing asthmatic symptoms (i.e., to reduce the severity, intensity, and/or duration of such symptoms). In such cases, the compositions are administered to an individual after asthmatic symptoms have developed.

Alternatively or additionally, the composition may be used to prevent or delay the onset of symptoms in an individual who has previously suffered asthmatic attacks, or to reduce the severity, intensity, or duration of subsequently-developed symptoms. Preferably, one or more antigens has been identified that is known to have induced, or at least to be correlated with, the onset of prior asthmatic attacks. In such cases, the inventive compositions are administered either prior to the onset of symptoms after a subsequent encounter with the antigen, or prior to the encounter.

The inventive compositions may also be administered prior to the development of asthmatic/allergic sensitivity to a particular antigen. Preferably, the compositions are administered substantially concurrently with exposure to an antigen that has not previously been associated with an asthmatic and/or allergic reaction in the individual. Without wishing to be bound by any particular theory, we propose that the inventive compositions may encourage the individual to adopt a Th1 response to the antigen. Given the mutually inhibitory aspects of Th1 and Th2 responses, the initial development of a Th1 response may inhibit, delay, or prevent subsequent Th1 reactions that could otherwise result in asthmatic and/or allergic symptoms.

Claim 1 of 6 Claims

What is claimed is:

1. A composition for treating allergies or asthma comprising effective amounts of aqueous extracts of Fructus Pruni Mume, Pericarpium Zanthoxyli Bungeanum, Herba cum Radice Asari, Rhizoma Coptidis, Corex Phellodendri, Rhizoma Zingiberis Officinalis, Radix Lateralis Aconiti Carmichaeli Praeparata, Ramulus Cinnamomi Cassiae, Radix Ginseng, and Radix Angelicae Sinensis.

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