KR101418458B1 - Composition comprising isozaluzanin C for prevention or treatment of allergy - Google Patents

Abstract

The present invention relates to a composition for the prevention or treatment of an allergic or inflammatory disease comprising isosalazin C,

Description

[0001] The present invention relates to a composition for preventing or treating an allergic or inflammatory disease, which comprises isozaluzanin C,

       The present invention is derived from research carried out by the Ministry of Education, Science and Technology / Korea Research Foundation as part of the general researcher support project.

[Task No .: 2011-0027501, Title of the Study: Action mechanism of atopy inhibitory substances involved in the regulation of the activity of interleukin-induced signaling molecules]

The present invention relates to a composition for preventing or treating an allergic or inflammatory disease, comprising isozaluzanin C.

The incidence of hypersensitivity or inflammatory immune diseases has been greatly increased with changes in the living environment and pollutants due to industrialization. Such irritable or inflammatory diseases have become one of the most common chronic diseases of modern people. Hypersensitivity or inflammatory immune disease is a progressive immune response in which the balance of immune tolerance and immune response is destroyed as in many other immune diseases. In addition, various factors have been reported to be associated with the onset and progression of the disease. Hypersensitivity or inflammatory immune disease refers to a case in which a defensive mechanism acting to protect a living body causes a disadvantageous action to the living body and causes allergy and inflammatory diseases.

Among them, the allergic reaction is initiated by the inflow of the antigen into the living body. The influenza antigens are recognized by antigen presenting cells, and T helper cells (Th) are differentiated into Th1 (T helper 1) cells and Th2 (T helper 2) cells. Cells differentiated by antigen stimulation become Th2 cells, leading to an imbalance between Th1 and Th2 responses in the body, resulting in allergic diseases and consequent allergic inflammation. Th1-type cytokines such as interferon-gamma may be used as an allergic disease inhibitor to balance the immune response (Whiteman SC, Spiteri MA. (2008), Journal of Inflammation-London 5: 8).

Allergies can be classified into five broad categories. (Immunoreactive hemolytic anemia), type 3: immunocomplex form (glomerulonephritis), type 4: cellular immunity form (tuberculin reaction (anaphylactic shock) ), And the fifth mode: irritation reaction (Grays' disease).

Among them, the first type of allergic reaction is a reaction induced by the interaction of IgE specific for an antigen and an antigen, and it belongs to various allergic diseases including atopy. Especially, mast cells are involved in allergic diseases and inflammatory diseases. Mast cells are present in the vascular tissues of mucosal surfaces of the intestines and skin, secrete inflammatory mediators and cytokines, and influence cell tracing (Beaven and Metzger (1993) Immunol Today 14 (5): 222-226; Bochner and Schleimer (2001) Immunol Rev 179: 5-15). IL-4 (Interleukin-4) induces the conversion of B cells into IgE and is involved in allergic reactions as it is overproduced. It infiltrates inflammatory cells and causes late stage allergic and inflammatory reactions (Metcalfe, Baram et al. 1997) Physiol Rev 77 (4): 1033-1079).

IL-5 is another major inflammatory inducer (Hamelmann, Wahn et al., (1999), Int Arch Allergy Immunol 118 (2-4): 90-94) When mast cells are stimulated in antigen-IgE complex, they are secreted from Th2 cells and increase eosinophil inflammation and mucus.

The present inventors conducted a study on a therapeutic agent derived from a natural substance that can effectively and safely administer an allergic or inflammatory disease, and found that isozaluzanin C extracted from laurel water is β-hexosus amylase in RBL-2H3 cells 5-dependent proliferation of Y16 early B cells by suppressing allergic and inflammatory-inducing and mediating microparticle secretion such as? -Hexosaminidase and IL-4, thus completing the present invention Respectively.

It is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of an allergic or inflammatory disease.

It is another object of the present invention to provide a method for producing isozaluzanin C from a natural substance.

One aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of an allergic or inflammatory disease comprising isosalujanin C of the formula:

      The isozaluzanin C contained in the composition of the present invention is a kind of sesquiterpene lactone compound and widely distributed in various plants. Especially, sesquiterpene lactone having a lactone structure is one of a group of plant natural compounds reported to have various physiological functions. This compound group has been known to have antitumor activity, cytotoxicity, antibacterial action, and insecticidal action, and pharmacological activities such as protection function and analgesic action of gastrointestinal cells have been reported.

The term "allergic disease" or "allergy ", as used herein, refers to various diseases, conditions, or abnormal conditions caused by hypersensitivity to certain substances in the human body, i.e., excessive reaction of the immune system against exogenous substances. In particular, an inflammatory mediator such as histamine is liberated from a substance introduced from the outside, thereby causing diseases.

In one embodiment of the invention, the allergic disease or allergy may be an allergic disease mediated by IgE (type 1 allergic reaction).

     In one embodiment of the invention, the inflammatory disease may be an inflammatory disease caused by secretion of inflammatory mediators of mast cells and secretion of cytokines mediated by inflammation and consequent proliferation of B cells.

 In one embodiment of the invention, the allergy can be selected from the group consisting of bronchial asthma, allergic rhinitis, atopic dermatitis, allergic conjunctivitis, allergic otitis media, urticaria and anaphylactic shock.

In one embodiment of the invention, isozaluzanin C is laureth nobilis ) leaves.

       In one embodiment of the invention, isozaluzanine C is

Drying the medicinal laurel leaf, warming the aqueous alcohol to 40 to 50 ° C, and then filtering to obtain an alcoholic extract,

       Concentrating the alcoholic extract under reduced pressure, adding chloroform and extracting it to obtain a chloroform layer fraction,

       Concentrating the chloroform layer fraction under reduced pressure, and separating by HPLC.

       In one embodiment of the invention, the water-soluble alcohol used for the extraction of laurel leaves may be methanol or ethanol.

The pharmaceutical composition of the present invention may comprise a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier to be contained in the pharmaceutical composition of the present invention is one which is usually used for the formulation and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, laurylvinyl pyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, But is not limited thereto. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Carriers and additives allowed as appropriate pharmaceutical is described in detail in ^{Remington's Pharmaceutical Sciences (19 th ed} ., 1995).

       The pharmaceutical composition of the present invention may be administered orally or parenterally, for example, by intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, or local administration.

       The appropriate dosage of the pharmaceutical composition of the present invention may vary depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate, . The dosage of the pharmaceutical composition of the present invention may preferably be 0.01-100 mg / kg (body weight) per day.

       The pharmaceutical composition of the present invention may be formulated into a unit dosage form by using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by those having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The pharmaceutical composition of the present invention may be formulated into pharmaceutical preparations including oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations such as ointments and creams, suppositories and sterilized injection solutions, Formulation, and may additionally comprise a dispersing or stabilizing agent.

In one embodiment of the invention, the pharmaceutical composition of the present invention may be used as a food composition. In this case, it further includes components which are usually added during the manufacture of food, and may include, for example, proteins, carbohydrates, fats, nutrients, flavoring agents and flavoring agents.

      Hereinafter, the present invention will be described in more detail with reference to specific examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

The composition comprising isosalzanin C, which is a sesquiterpene lactone compound according to one embodiment of the present invention, inhibits the expression of allergenic microparticles and the expression of inflammatory cytokines and inhibits cytokine mediated cell proliferation, Can be usefully used for the prevention or treatment of inflammatory diseases.

Fig. 1 shows the cytotoxic effect of isozaluzan C on RBL-2H3 cells according to one embodiment of the present invention.
Figure 2 shows the inhibitory effect of isozaluzanin C on the release of beta-hexosaminidase and the positive control ketotifen in RBL-2H3 cells.
Figure 3 shows isozaluzanin C for IL-4 release in RBL-2H3 and the inhibitory effect of the positive control, ketotifen.
FIG. 4 is a graph showing the results of RT-PCR showing the inhibitory effect of isozaluzan C and the positive control, ketotifen on the expression of IL-4 mRNA in RBL-2H3 cells.
FIG. 5 is a graph showing Western blotting results showing quantitation of IL-4 protein expression in RBL-2H3 cells and isozylazinin C and a positive control, ketotifen, and quantification thereof.
Figure 6 shows the inhibitory effect of isosaluzanin C and a positive control, tyrphostin AG490, on IL-5 dependent Y16 cell proliferation.

Example  One. Isozaluzanine  Preparation of C compounds

Isozaluzanin C was isolated from laurel leaves. First, 20 kg of 100% ethanol was added 6 times to 10 kg of laver laurel leaf after shade drying, and extracted at 55 ° C for 48 hours to extract 2.4 kg of ethanol layer. The 100% ethanol extract layer obtained above was concentrated under reduced pressure, 20 L of chloroform was added six times, and the chloroform layer was fractionated to obtain 282.79 g of a chloroform layer fraction.

       The chloroform layer fractions of the laurel leaves obtained above were separated by chromatography on a silica gel column (YMC-Pack SIL), and the fraction was monitored by TLC while using a mixed solvent of hexane and ethyl acetate, a gradient of acetonitrile and methanol 50% ethyl acetate / 50% hexane, 60% ethyl acetate / 10% ethyl acetate / 90% hexane, 20% ethyl acetate / 80% hexane, 30% ethyl acetate / 70% / 40% hexane, 70% ethyl acetate / 30% hexane, 100% ethyl acetate, 100% acetonitrile, 100% methanol) to give 11 fractions.

The fraction 4 obtained above was separated by HPLC using a normal silica column (HPLC: YMC-silica column, 25 x 1 cm), eluted with a mixed solvent of ethyl acetate and hexane (12% ethyl acetate / hexane) Pure fractions of isozaluzan C were obtained. The isolated compounds were identified by comparison with data published via ¹ H-NMR, ¹³ C-NMR, COZY and HSQC (Bohlmann F, Brindopke G, et al. (1978): Phytochemistry 1978: 17: 475-482).

Example  2. Isozaluzanine  Anti-allergic effect of C

In this Example, the anti-allergic effect of isozaluzan C obtained in Example 1 was confirmed.

Statistical significance was determined by t-test (Sigma Plot 10.0 software, SPSS, USA) and p <0.05 was considered statistically significant .

2-1. RBL -2 H3  Cell culture and cytotoxicity

RBL-2H3 cells are very similar to basophilic and mast cells, express IL-4 rapidly, and are most conveniently used in the study of the microparticle secretion of mast cells. On the surface of these cells, there is a large amount of receptor FcεRI, which has a strong binding force to IgE, and such IgE-FcεRI binding is induced by a large amount of antigen. (Barsumian, Isersky et al., (1981), Eur J Immunol 11 (4): 317-323 ), serotonin (Taurog, Fewtrell et al., 1979) from cells by the reaction of RBL- , J Immunol 122 (6): 2150-2153 ) and beta-hexosaminidase (Schwartz, Austen et al. (1979), J Immunol 123 (4): 1445-1450) Mediates are released. These substances play a major role in causing allergic symptoms such as vasodilation, mucus secretion, and bronchodilation.

To investigate the antiallergic effect of isozaluzan C, RBL-2H3 cells (ATCC No. CRL-2256) were cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS) (Gibco-BRL, USA) and antibiotic solution (Sigma-Aldrich, USA) supplemented with DMEM (Sigma-Aldrich, A5955, USA) at 37 ° C and 5% CO ₂ .

The cytotoxicity of isozaluzan C was tested against RBL-2H3 cells. MTT assay was used to confirm cell viability. RBL-2H3 cells (5 x 10 ⁵ cells / mL) were inoculated into 100 μl / well of culture medium in 96-well plates and cultured under the conditions described above. After incubation for 24 hours, the cells were reacted with various concentrations of isozaluzanin C for 4 hours and then MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide) (250 [mu] g / ml) were added and cultured at 37 [deg.] C for 3 hours. Then 100 μl of sodium dodecyl sulfate (SDS) solution (50% DMF and 20% SDS in water) was added per well to dissolve the formazan crystals. Cell viability was measured by absorbance at 595 nm, and the survival rate of the control group without isozulozanin C was 100%.

Figure 1 shows the effect of isozaluzan C on survival of RBL-2H3 cells. The survival rate of RBL-2H3 cells was 95% when compared with the control group at 0.32 μM to 80 μM.

2-2. RBL -2 H3  The beta - Hexosaminidase  Secretion inhibition

To measure allergen-induced microparticle secretion in RBL-2H3 cells, sensitization with IgE was followed by an immune response with DNP-BSA. The amount of the fine particles secreted was measured by beta-hexosaminidase secretion.

RBL-2H3 cells (5 x 10 ⁴ cells / mL) were inoculated into 100 μl / well of culture medium in 96-well plates and cultured under the conditions described in 2-1. At the time of culture, the cells were sensitized with final 1 / / mL of anti-DNP-IgE (Sigma-Aldrich, USA). After incubation, the obtained cells were washed twice with HEPES buffer to remove the remaining IgE without binding to the cells. Cells were then treated with isozaluzan C and 45 μl of a releasing mixture (116.9 mM NaCl, 5.4 mM KCl, 0.8 mM MgSO ₄ .7H ₂ O, 5.6 mM glucose, 25 mM HEPES, 2.0 mM CaCl 2 ₂ and 1.0 mg / mL BSA at pH 7.7) for 20 minutes. Then, 5 μl of DNP-BSA (4 μg / mL) dissolved in PBS was added to the cells and reacted for 1 hour. After the reaction, only the supernatant was removed from the plate and transferred to a new 96-well plate and 50 μl of a substrate solution (5 mM 4-nitrophenyl N-acetyl-β-D-glucosamid in 50 mM citrate buffer, pH 4.5) Followed by incubation at 37 DEG C for 90 minutes. Termination buffer _{(stopping buffer, 0.5 M NaCO 3} / NaHCO 3, pH 10.0) and the reaction was terminated by the addition of 50 ㎕. Absorbance was measured at 405 nM in a microplate reader.

Negative controls were not treated with isozaluzan-C, and positive controls were treated with ketotifen. Ketotifen is an inhibitor of IgE-mediated degranulation and is frequently used with RBL-2H3 cells for immune and allergy studies. Ketotifen is known to selectively inhibit histamine release in tissue-bound mast cells and inhibit histamine release.

The release inhibition (%) by isozaluzan-C was calculated by the following equation.

Suppression (%) = [1- (T-C) / (I-C)] x 100

(+), DNP-IgE (+), DNP-BSA (-), isozaluzanin-

Test (T): Cell (+), DNP-IgE (+), DNP-BSA (+), isosaluzanin-

(+), DNP-IgE (+), DNP-BSA (+), isozaluzanine-

Upon isozaluzanin C treatment, the amount of beta -hexosaminididase secreted in RBL-2H3 cells was decreased in a concentration-dependent manner. Figure 2 shows the effect of isozaluzanin C compound on beta -hexosaminidase secretion with positive control ketotifen. It was confirmed that isozaluzanin C decreased the amounts of beta -hexosaminidase secretion by 0.5%, 17.2%, and 58.5% at the respective concentrations (1.6, 8, and 40 μM), respectively. (IC ₅₀ = 29.6 [mu] M).

2-3. ELISA Using RBL -2 H3  Cell IL -4 measurement of secretion

RBL-2H3 cells (5 x 10 ⁵ cells / mL) were inoculated into 100 μl / well of culture medium in 96-well plates and cultured under the conditions described in 2-1. At the time of culture, the cells were sensitized with final 1 / / mL of anti-DNP-IgE (Sigma-Aldrich, USA). After incubation for 24 hours, isozaluzanin C was treated at different concentrations and incubated for 3 hours, followed by treatment with DNP-BSA and further incubation for 2 hours. The amount of the end-acquired cell Rat IL-4 ^TM Platinum ELISA kit USA) IL-4 secretion, as a manual for the companies use were measured. The results showed that treatment with isozaluzanin C decreased IL-4 secretion in a concentration-dependent manner and is shown in Fig. Figure 3 shows the effect of isozaluzanin C compound on IL-4 secretion versus positive control ketotifen. Isozaluzanine C reduced IL-4 secretion by 0.1%, 20.2%, and 48.0% at the respective concentrations (1.6, 8, and 40 μM), respectively. (IC ₅₀ = 44.1 [mu] M).

2-4. Using polymerase chain reaction IL -4 mRNA  Measurement of expression level

RBL-2H3 cells (5 x 10 ⁵ cells / mL) were inoculated into a culture medium of 1000 μl / well in a 12-well plate and cultured under the conditions described in 2-1. At the time of culture, the cells were sensitized with final 1 / / mL of anti-DNP-IgE (Sigma-Aldrich, USA). After culturing for 24 hours, isozaluzan C was treated at different concentrations, incubated for 3 hours, treated with DNP-BSA, and further incubated for 1 hour. After the cultured cells were recovered by centrifugation, total RNA was extracted using an easy-BLUE ™ Total RNA Extraction Kit (Intron Biotechnology, Korea). CDNA was synthesized using quantitative RNA and ONE-STEP RT-PCR premix kit (Intron Biotechnology, Korea).

The polymerase chain reaction was carried out in a GeneAmp PCR System 9700 (Applied Biosystems) using the cDNA obtained above as a template, denaturation at 94 DEG C for 30 seconds using the primer described below, annealing at 57 DEG C for 23 seconds, and annealing at 35 DEG C for 72 DEG C Lt; RTI ID = 0.0 &gt; 35 &lt; / RTI &gt; times. Beta -actin was used as an internal control.

IL-4:

The forward primer 5'-GGGTGCTTCGCAAATTTACTT-3 '(SEQ ID NO: 1)

The reverse primer 5'-ACCGAGAACCCCAGACTTGTT-3 '(SEQ ID NO: 2)

beta -actin:

The forward primer 5'-TCTGTGTGGATTGGTGGCTCTA-3 '(SEQ ID NO: 3)

The reverse primer 5'-CTGCTTGCTGATCCACATCTG-3 '(SEQ ID NO: 4)

Figure 4 shows the effect of isozaluzanin C compound on the amount of IL-4 mRNA expression versus the positive control ketotifen. Isozaluzanine C decreased IL-4 mRNA expression levels by 0.0%, 12.2%, and 51.5% at the respective concentrations (1.6, 8, and 40 μM), respectively (IC ₅₀ = 34.6 [mu] M).

2-5 Western blotting  Used IL -4 Protein expression level measurement

RBL-2H3 cells (5 x 10 ⁵ cells / mL) were inoculated into 2000 μl / well of culture medium in 6-well plates and cultured under the conditions described above. Upon incubation, cells were sensitized with anti-DNP-IgE at a final concentration of 1 / / mL. After culturing for 24 hours, isozaluzan C was treated at different concentrations, incubated for 3 hours, treated with DNP-BSA, and further incubated for 1 hour. The cultured cells were recovered by centrifugation, and total cellular proteins were extracted using PRO-PREP protein solution (Intron biotechnology, Korea). Then, the protein concentration was measured using a protein assay solution (Intron biotechnology, Korea: Pro-Measure ^TM ). 20 μg of the cell protein was electrophoresed on a SDS-PAGE gel and transferred to a PVDF membrane (Amersham Pharmacia Biotech, UK). The membrane was incubated with TBST (5% skim milk in tris buffered-0.1% tween 20) Blocking. Then, the cells were reacted with primary antibody (IL-4 antibody, Santa Cruz biotechnology, USA) for 24 hours at 4 ° C, washed with TBST, and incubated with secondary antibody (biotinylated goat anti- mouse antibody, Santa Cruz biotechnology, USA) For 1 hour. After washing, the image was analyzed with an image analyzer LAS1000 (Fuji photo film, Japan). All western blotting analyzes were repeated at least three times.

Figure 5 shows Western blotting results showing the concentration-dependent inhibitory effect of isozaluzanin C and positive control ketotifen on the expression of IL-4 protein and a graphical representation thereof. Iso jalu mezzanine C had IL-4 reduced the protein expression level of each of 5%, 17%, and 62% at each concentration (1.6, 8, and 40 μM) (IC ₅₀ = 31.4 [mu] M).

2-6. IL -5 dependency Y16 early  Inhibition of B cell proliferation

Y16 cells are B cells in their initial state and proliferate according to the presence of IL-5. In this example, the proliferation of IL-5 dependent Y16 cells was measured to examine the effect of isozaluzan C on IL-5 activity.

Y16 cells (provided by Professor K. Takatsu of Tokyo University) were cultured in RPMI (10.4 mg / mL RPMI-1640, 24 mM NaHCO _{3 ,} 100x antibiotic solution (A5955, Sigma-Aldrich, USA), 50 μM 2-mercaptoethanol) At a final concentration of 10 U / mL mIL-5, and cultured at 37 ° C in 5% CO ₂ . After culturing, the culture was centrifuged at 1000 rpm for 10 minutes to collect the cells. The cells were suspended in the culture solution and dispensed into a 96-well microplate in an amount of 100 5 each at 5 × 10 ⁴ cells / well. Of isoleucanin C in mIL-5 and 10% DMSO. In the control group, only 10% DMSO was added instead of isozaluzanine C, and the Inducer group was treated with 10% DMSO and mIL-5 in place of isozaluzanin C. As a positive control, tyrphostin AG490 inhibiting IL-5-mediated cell proliferation was used. Cells treated with the sample were incubated at 37 ° C in 5% CO ₂ for 48 hours and then incubated with MTT (3- (4,5-dimethylthiazol-2yl) -2,5-diphenyltetrazolium bromide) (250 μg / . After the reaction, sodium dodecyl sulfate (SDS) (50% DMF, 20% SDS in water) was added at a rate of 100 μl per well, followed by reaction for 24 hours. Survival of Y16 cells was measured at 540 nm ( A ₅₄₀ ) using a microplate reader (Molecular Device, USA).

The inhibitory effect on IL-5 was calculated according to the following equation:

Inhibition (%) = [1- (sample A ₅₄₀ - control A ₅₄₀ ) / (derivative A ₅₄₀ - control A ₅₄₀ )] x 100

All experiments were performed with a treatment concentration of isozaluzan C ranging from 0.8 μM to 40 μM. FIG. 6 shows the cell proliferation inhibition efficiency (%) of isozaluzan C when the cell growth rate of the control group is taken as 100% as compared with the standard control substance, Tirfostin AG490. Iso jalu mezzanine C had 10.4%, respectively, the Y16 cell proliferation at each concentration (0.8, 4, 20, and 40 μM), 27.5%, decreased 34.0%, and 56.2% (IC ₅₀ = 39.4 [mu] M).

<110> Seoul National University R & DB Foundation <120> Composition containing isozaluzanin C for prevention          or treatment of allergy or inflammatory disorders <160> 4 <170> Kopatentin 1.71 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> forward primer for IL-4 <400> 1 GGGTGCTTCGC AAATTTTACT T 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer for IL-4 <400> 2 ACCGAGAACC CCAGACTTGT T 21 <210> 3 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> forward primer for beta-actin <400> 3 TCTGTGTGGA TTGGTGGCTC TA 22 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> reverse primer for beta-actin <400> 4 CTGCTTGCTG ATCCACATCT G 21

Claims (3)

A composition for preventing or treating allergies comprising isozaluzanin C as an active ingredient. The composition of claim 1, wherein said allergy is selected from the group consisting of bronchial asthma, allergic rhinitis, atopic dermatitis, allergic conjunctivitis, allergic otitis media, urticaria, asthma and anaphylactic shock. 2. The composition of claim 1, wherein said isozaluzanin C is derived from an extract of Laurus nobilis leaf.

Images