HK1084953A - Synthetic lactone formulations and method of use - Google Patents

Description

Synthetic lactone formulations and methods of use thereof

Technical Field

This application claims priority to the submission of U.S. provisional patent application 60/424,045 on 5/11/2002.

The present invention is in the field of pharmaceutically active lactones, pharmaceutical formulations thereof, and methods of use thereof, as well as methods of synthesizing chemically functional lactones useful as anti-cancer, anti-infective, and anti-inflammatory agents.

Background

Although a variety of different compounds have been developed that are effective in treating infections, cancers, and other disorders, there remains a need to develop new compounds that are effective, more selective, and more effective at lower doses, have fewer side effects, or can treat diseases or disorders that are resistant to treatment with known compounds.

Chemotherapeutic agents are used to treat infections, cancers, abnormal proliferative disorders (endometriosis, restenosis, psoriasis), and other disorders. Most chemotherapeutic agents have side effects due to lack of specificity. For example, cancer is one of the leading causes of death. One of the primary modes of treating cancer, chemotherapy, is used to specifically limit cell growth and proliferation. Most chemotherapeutic agents also affect tumors and rapidly proliferating cells in normal tissues (e.g., bone marrow, hair follicles, etc.) and produce several side effects including alopecia, nausea, vomiting, and bone marrow function suppression. In addition, the effectiveness of these formulations often diminishes over time due to the development of resistance.

Resistance to chemotherapeutic agents is more pronounced in the treatment of bacterial or fungal diseases. For example, Helicobacter pylori (Helicobacter pylori) causes gastric disorders in a large population in the united states. Lack of effective treatment for these disorders can lead to the development of peptic ulcers, gastritis, dyspepsia and gastric cancer. Another common bacterial disease is periodontal disease, the major cause of disease being bacterial plaque, which can lead to the development of periodontitis and ultimately tooth loss.

It is therefore an object of the present invention to provide a new class of compounds which are effective as anti-infective, anti-proliferative and anti-inflammatory agents.

It is another object of the present invention to provide an effective anti-tumor agent having specific cytotoxicity to minimize side effects.

It is another object of the present invention to provide anti-infective agents that are specific and different from many other drugs currently in use, providing an alternative treatment for drug resistant organisms.

Summary of The Invention

Natural and synthetic compounds of formula Ia, Ib, and Ic having a lactone structure, and methods of using and making the compounds, as well as compositions for the administration of the compounds, have been developed. The compounds are useful as antibacterial, antifungal and anti-inflammatory agents, and are useful for treating proliferative disorders such as melanoma, leukemia, breast cancer, lung cancer, ovarian cancer, colon cancer, esophageal cancer, liver cancer and lymphatic cancer. The compounds are also useful for treating or preventing inflammatory diseases such as artherosclerosis, pulmonary fibrosis, systemic lupus erythematosus, pancreatitis, sarcoidosis, glomeruloitis and organ transplant rejection. In addition, they are also effective in the treatment or prevention of bacterial and fungal infections, including the treatment of peptic ulcers, gastritis, dyspepsia and gastric cancer, gingivitis and periodontitis.

A process for preparing compounds of formula Ia, Ib and Ic comprising: a) providing a precursor having a lactone structure, and b) reacting the precursor with one or more chemical reagents to produce the compound. The compounds may be further derivatized by reaction with nucleophiles such as alcohols, alkoxides, amines, or any other neutral or negatively charged nucleophile.

Detailed Description

I. Lactone compositions

Lactones

Lactones and their derivatives each bearing a hydroxyl group in the gamma position are disclosed. Lactones and derivatives thereof can be synthesized or obtained from natural sources. In one embodiment, the lactones and derivatives can be isolated chromatographically from plants having the taxonomic name Securidaca virgata, belonging to the Polygalaceae family of plants. The term "lactone" as used herein includes any organic compound having a five-membered ring lactone structure in which the oxygen atom of the C ═ O group can be substituted with a sulfur atom or a nitrogen group. The term "derivative" as used herein refers to any compound made by reacting a lactone with one or more chemical reagents. The term also refers to any product formed by ring opening of a lactone with an organic or inorganic nucleophile, such as an acid, ester, amide, or any other product thereof.

In one embodiment, the lactone has the following chemical structure:

formula Ia

Wherein R is₁-R₆Independently a hydrogen atom, a halogen atom, a hydroxyl group or any other organic group containing any number of carbon atoms, preferably 1 to 8 carbon atoms, and optionally including heteroatoms such as oxygen, sulfur or nitrogen groups, in linear, branched or cyclic structures, representative of R₁-R₆The radicals being H, alkyl, substituted alkyl, allylSubstituted allyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, alkoxy, substituted alkoxy, alloxy, phenoxy, substituted phenoxy, aryloxy, substituted aryloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C₁-C₂₀Cyclic, substituted C₁-C₂₀A ring, heterocycle, substituted heterocycle, amino acid, peptide, or polypeptide group;

z is a heteroatom selected from the group consisting of oxygen, sulfur or nitrogen radicals in a linear, branched or cyclic form; and

x is a heteroatom selected from the group consisting of oxygen, sulfur or nitrogen radicals in a linear, branched or cyclic form.

In another embodiment, the lactone has the following chemical structure:

formula Ib

Wherein R is₁-R₄Independently a hydrogen atom, a halogen atom, a hydroxyl group or any other organic group containing any number of carbon atoms, preferably 1 to 8 carbon atoms, and optionally including heteroatoms such as oxygen, sulfur or nitrogen groups, in linear, branched or cyclic structures, representative of R₁-R₄Is H, alkyl, substituted alkyl, allyl, substituted allyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynylPhenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, alkoxy, substituted alkoxy, alloxy, phenoxy, substituted phenoxy, aryloxy, substituted aryloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C₁-C₂₀Cyclic, substituted C₁-C₂₀A ring, heterocycle, substituted heterocycle, amino acid, peptide, or polypeptide group;

x is a heteroatom such as oxygen, sulfur or nitrogen group in a linear, branched or cyclic structural formula; and

z is a heteroatom such as an oxygen atom, sulfur atom or nitrogen group in a linear, branched or cyclic structural formula.

In another embodiment, the lactone with an alpha methylene group has the following structure:

formula Ic

Wherein R is₁-R₄Each of which may be a hydrogen atom, a halogen atom, a hydroxyl group or any organic group containing any number of carbon atoms, preferably 1 to 8 carbon atoms, and optionally including in a linear, branched or cyclic structure a heteroatom such as an oxygen, sulfur or nitrogen group, representative R₁-R₄Is alkyl, allyl, substituted alkyl, alkenyl, allyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted aryl, substituted heteroaryl,halogen, hydroxy, alkoxy, substituted alkoxy, alloxy, phenoxy, substituted phenoxy, aryloxy, substituted aryloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C₁-C₂₀Cyclic, substituted C₁-C₂₀A ring, heterocycle, substituted heterocycle, amino acid, peptide, or polypeptide group;

z is a heteroatom selected from the group consisting of oxygen, sulfur or nitrogen radicals in a linear, branched or cyclic form; and

x is a heteroatom selected from the group consisting of oxygen, sulfur or nitrogen radicals in a linear, branched or cyclic form.

Representative lactones of formulae Ia, Ib and Ic are listed in Table 1:

TABLE 1 representative synthetic endosomes

Pharmaceutically acceptable acid addition salts of the compounds of formula Ia, Ib or Ic can be prepared by conventional methods, i.e. by treating a solution or suspension of the free base of formula I with about one stoichiometric equivalent of the drug and the acid. Conventional concentration and recrystallization techniques are used to isolate the salt.

Pharmaceutically acceptable base addition salts of compounds of formula 1 containing an acid group may be prepared from the acid by conventional methods, for example by reaction with about one stoichiometric equivalent of a base.

B. Excipient

Lactones and functional derivatives can be prepared for enteral, parenteral or topical administration using conventional techniques (see, e.g., Encyclopedia of controlled Drug Delivery, Edith mathiow, ed., John Wiley & Sons, inc., New York, 1999). Effective dosages can be determined based on in vitro assays well known to those skilled in the art, such as the assays described in the examples.

Suitable pharmaceutical carriers for parenteral delivery include sterile saline, phosphate buffered saline, pyrogen free sterile media and standard microparticle formulations for injection, including polymeric microspheres, microcapsules, liposomes and emulsions. These may include degradable polymers such as polylactic acid and polyglycolic acid, and copolymers thereof, polyanhydrides, polyorthoesters, polyhydroxyalkanoates.

Suitable pharmaceutical carriers include talc, acacia, lactose, starch, magnesium stearate, cocoa butter, aqueous or non-aqueous vehicles, fatty substances of animal or vegetable origin, paraffin derivatives, glycols, various wetting, dispersing or emulsifying agents and preservatives.

For injection, the lactone is typically prepared as a solution or suspension in a liquid carrier.

For topical administration, the lactone may be formulated as an ointment, cream, lotion, gel, spray, or controlled or sustained release formulation (e.g., transdermal patch).

For enteral administration, the lactone may be formulated as a tablet, capsule, granule, suppository, suspension or solution, dissolved or encapsulated in a sugar such as lactose, an inert compound such as magnesium stearate, a paraffin derivative, ethylene glycol or acacia, and excipients. The formulation may further include dyes, flavourings, preservatives, dispersing or emulsifying agents, or substances which modify the release or stability properties of the formulation.

These active compounds may be used in combination with another pharmaceutically acceptable antimicrobial agent: nitroimidazole antibiotics, for example, tinidazole and metronidazole; tetracyclines, such as tetracycline, doxycycline and minocycline; penicillins, such as amoxicillin and mezlocillin; cephalosporins, such as cefaclor, cefadroxil, cephaeline, cefuroxime axetil, cefonicid, cefpodoxime proxetil, ceftazidime and ceftriaxone; carbapenems, such as imipenem and minoxidil; aminoglycosides, such as paromomycin; macrolide antibiotics such as erythromycin, methylerythromycin and azithromycin; lincosamide antibiotics, such as clindamycin; rifamycins, such as rifampin; and nitrofurantoin.

The combination of these compounds with a pharmaceutical acid-lowering agent can be used to treat acid-related disorders, such as acid pump inhibitors, e.g., omeprazole and lansoprazole, or H₂Antagonists such as ranitidine, cimetidine and famotidine.

Synthesis of lactones

The syntheses of formulae Ia, Ib and Ic include: a) forming an intermediate or precursor having a lactone structure, and b) reacting the intermediate with one or more suitable chemical reagents to produce lactones of general formulae Ia, Ib and Ic.

In one embodiment, the method comprises: a) providing a precursor having the following structure

Formula II

And b) reacting the precursor with one or more suitable chemical reagents to provide lactones of general formulae Ia, Ib and Ic (scheme 1).

(reaction scheme 1)

As shown in the reaction scheme 1, acetylene can be phosphine with acetaldehyde in the presence of [ CH ]₃(CH₂)₃]₃P to produce compound A, which undergoes a ring recombination reaction to form compound B in its enol form. The enolic compound B is in equilibrium with its ketone-form compound C. Compounds B or C form compounds E (formula Ic) or F (formula Ia, Ib) in the presence of a base such as butyllithium, sodium carbonate, sodium hydroxide or sodium methoxide or ethoxide. Alternatively, the enol compound B can be reacted with NaBH₄Reduction is carried out to form a saturated compound D. Reaction of Compound D with HCO₂Concentration of Et reacts to form the cyclic enol ester, compound G, which is then reduced by formylating the aldehyde to form compound H. Compound H can be readily derivatized to form compound I (formula Ia) using, for example, haloalkyl in the presence of a base such as sodium carbonate, sodium hydroxide or sodium methoxide or ethoxide.

More functionalized lactones can be prepared by synthetic methods readily available in the art (see, e.g., March, Advanced organic chemistry, fourth edition, 1992, Wiley-Interscience Publication, New York).

Pharmaceutically acceptable salts of the lactone compounds of formula Ia-c, in the form of an acid or a base such as an amine, may be prepared in conventional manner by treating a solution or suspension of the compound of formula Ia-c with about one chemical equivalent of a pharmaceutically acceptable base or acid. Conventional concentration and recrystallization techniques are used to isolate the salt.

Methods of treatment

A. Disorder to be treated

Lactones are useful as anti-infective, anti-proliferative and anti-inflammatory agents to prevent or treat a broad spectrum of disorders. In particular, the lactones are useful in the treatment of disorders including, for example, cancer, gingivitis, periodontitis, helicobacter pylori-induced diseases, bacterial-induced diseases, fungal-induced diseases, and inflammatory diseases.

The lactones can be prepared into fungicidal compositions, antibacterial compositions, anticancer compositions, and anti-inflammatory compositions. Fungicidal compositions comprise a fungicidally effective amount of a compound of formula Ia, Ib or Ic or a salt thereof and an inert pharmaceutical carrier. The fungicidal compositions are particularly effective against Saccharomyces cerevisiae (Saccharomyces cerevisiae), Candida albicans (Candida albicans) and other Candida species such as Candida glabrata (Candida glabrata), Candida krusei (krusei), Candida tropicalis (tropicalis), Candida pseudothermalis (pseudotropicalis) and Candida parapsilosis (parapsilosis), Aspergillus fumigatus (Aspergillus fumigatus), Aspergillus flavus (Aspergillus flavus), Aspergillus niger (Aspergillus niger), Cryptococcus neoformans (Cryptococcus eodormans), Microsporum canis (Microsporium canis), Trichosporon rubrum (Trichophyton rubrum), Trichosporon mentagrophytes (Trichophyton trichoderma) and Trichosporon trichoderma (Trichosporon), and particularly against invasive conditions of the respiratory, vaginal, and cutaneous tracts such as bronchogenic, pulmonary and pulmonary mucormycosis, as pulmonary cryptophytes, pulmonary and pulmonary diseases. The composition can also be used for preventing fungal infections of the innate and adaptive immune suppression systems.

The lactones can be administered to treat or prevent inflammatory diseases. Representative inflammatory diseases include artherosclerosis, pulmonary fibrosis, systemic lupus erythematosus, pancreatitis, sarcoidosis, glomerulonephritis, and organ suppression rejection, such as kidney transplantation, liver transplantation, lung transplantation, and heart transplantation.

The lactones can also be administered to treat proliferative disorders, including cancer. Representative types of cancers that exhibit inhibitory effects on cell growth or proliferation include melanoma, leukemia, breast cancer, lung cancer, ovarian cancer, colon cancer, esophageal cancer, liver cancer, and lymphoma. Other types of abnormal proliferative disorders that lactones may be effective in treating include endometriosis and restenosis resulting from abnormal hyperproliferation of endothelial tissue following angioplasty.

The antibacterial composition may be administered to treat or prevent bacterial disorders including, for example, diseases caused by helicobacter pylori such as gastric ulcer, gastritis, dyspepsia and gastric cancer. The antibacterial composition effective for the treatment and prevention of diseases caused by helicobacter pylori may be used in combination with another pharmaceutically acceptable antimicrobial agent or a pharmaceutically acceptable acid-lowering agent.

Lactone antibacterial compositions are effective in treating or preventing oral diseases such as periodontitis, plaque and gingivitis, the compositions are effective against specific anaerobic gram negative microorganisms associated with gingivitis. The lactone composition can be dissolved in an oral medium and topically applied to the oral cavity of an animal in the form of an oral formulation. The oral formulations may be in the form of, for example, mouthwashes and toothpastes. The lactone composition can also be prepared in an oral composition for improving oral hygiene by topically applying the lactone composition to the oral cavity.

B. Dosage form

The effective amount will depend on the disease or condition to be treated, the mode of administration and the dosage form. Effective dosages can be routinely determined based on those determined by in vitro assays described in the examples.

The high activity and low molecular weight of a class of alpha methyllactones, 4, 5-dihydro-3-methylene-2 [3H ] furanones ("Securolide" or "LMSV-6") against E.coli (Escherichia coli), Klebsiella pneumoniae (Klebsiella pneumoniae), Pseudomonas aeroginosa (Pseudomonas aeroginosa), Staphylococcus aureus (Staphylococcus aureus) are highly beneficial. Advantages of Securolide include its ability to facilitate pharmacological reactions quickly and conveniently; its ability to cross cell membrane barriers, where high molecular weight is the major obstacle, and its potential activity against pseudomonas, one of a number of drug resistant microorganisms.

A method of combating fungal infections comprises applying a fungicidally effective amount of a compound of formula I or an acid addition salt thereof, topically to the skin or mucosa by oral, rectal, parenteral or topical route, but preferably the route of administration is oral. Depending on the method of administration, the condition being treated and the particular compound, a daily dose of 1-5mg/kg is typical.

The compounds may be administered alone, but will generally be administered in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceutical practice. For example, they can be administered orally, or in the form of tablets containing excipients such as starch or lactose, or in the form of capsules, either alone or in admixture with excipients, or in the form of elixirs or suspensions containing flavoring or coloring agents. In the case of animals, they are preferably contained in the animal feed or drink in a concentration of about 5 to 5000ppm, preferably about 25 to 500 ppm. They may be injected parenterally, for example, intramuscularly, intravenously or subcutaneously. For parenteral administration, they are best used in the form of a sterile aqueous solution containing other solutes, for example, sufficient salts or glucose, to render the solution isotonic. In the case of animals, the compounds can be administered intramuscularly or subcutaneously at dosage levels of about 0.1 to about 50 mg/kg/day, preferably about 0.2 to 10 mg/kg/day in a single administration or in divided doses four times per day.

The compounds can be administered to humans by oral or parenteral routes for the treatment of helicobacter pylori infections, and can be administered in a single dose or in divided doses of about 0.1 to about 50mg/kg, preferably about 0.5 to 50 mg/kg/day, in four oral administrations. For intramuscular or intravenous administration, the dosage level is from about 0.1 to about 100 mg/kg/day, preferably from about 0.5 to about 50 mg/kg/day. While intramuscular administration may be in a single dose or divided into up to 4 doses, intravenous administration may include continuous instillation. Various modifications will be apparent to those skilled in the art depending upon the weight and disease state of the subject being treated and the particular route of administration chosen.

The second antimicrobial agent and acid-lowering agent may be administered in the same manner as the compounds of the invention discussed above. Thus, depending on the particular agent, administration may be orally at about 0.1 to about 500mg/kg, e.g., about 1 to 3g of the second antimicrobial agent per day, and about 40 to 80mg of the acid-lowering agent per day, or by injection at about 0.1 to about 200mg/kg per day.

C. Mode of administration

The lactone composition can be administered to the warm-blooded animal by any suitable means of administration. The mode of administration may be local administration or systemic administration. The mode of administration may vary depending on the disorder to be treated or prevented.

The composition can be administered to the animal by enteral, parenteral or topical administration. Representative modes of administration include: oral administration, nasal administration, pulmonary administration, vascular administration, subcutaneous injection, transdermal administration, mucosal administration and administration by the oral, rectal or vaginal route. The invention is further illustrated with reference to the following non-limiting examples.

Example 1: microbiological susceptibility assay

Culture medium:

solvent: phosphate buffer 0.1N, pH 8.0

Antibiotics: LMSV-6 (Securolide):

Securolide

coating the culture medium: 2mL/100mL medium

Inoculum: 4 mL/Petri dish

Preparation of the culture Medium

Culture medium of Staphylococcus aureus (USP 23<81>)

.

The

A

.

.

The

Adding water to about 1.0L

pH6.6 + -0.1 after sterilization

Culture medium of Pseudomonas aeroginosa (USP 23<81>)

The

.

.

The

.

.

Adding water to 1.0L

After sterilization, the pH value is 7.2-0.1

Culture medium of Escherichia coli and Klebsiella pneumoniae (K. Neumoniae) (according to USP 23<81>)

.

A

.

.

.

The

The

Adding water to 1.0L

pH7.0 after sterilization 0.05

Antibacterial Activity assay (minimum inhibitory concentration, MIC)

(MIC) detection No.1

The method comprises the following steps: mueller Hinton medium at pH 8 was poured into plates

Buffer pH 8 for ceftriaxone mode dilution

2% TWEENTM 20 was used for sample dilution

The microorganisms used were: sarcina lutea (Sarcina lutea)

Sample (S): LMSV-6(5 pure μ L applied to sensitive dishes)

Mode (P): ceftriazine (with 10 mug plate)

P(mm) S(mm)

12 40

1544X 127X 100 295.35% (considering 10. mu.g of ceftriaxone)

16 43

43 127

The sample zone of inhibition is three times larger than the mode zone.

(MIC) assay No 2

The method comprises the following steps: mueller Hinton medium at pH 8 was poured into plates

Buffer pH 8 for ceftriaxone dilution mode

2% TWEEN 20 was used for sample dilution

The microorganisms used were: sarcina lutea (berk.) Kuntze

Sample (S): 16.0 microliter/100 mL and apply 10. mu.L onto sensitive dishes)

Mode (P): ceftriazine (with 10 mug plate)

P(mm) S(mm)

18 20

16 23

15 19

49 62

X62 × 100 126.53% (considering 10 μ g of ceftriaxone)

49

(MIC) detection No.3

The method comprises the following steps: mueller Hinton medium at pH 8 was poured into plates

Buffer pH 8 for ceftriaxone dilution mode

2% TWEEN 20 was used as LMSV-6(Securolide) dilution

The microorganisms used were: sarcina lutea (berk.) Kuntze

Sample (S): (10. mu.l/10 mL and 20. mu.L applied to a sensitive plate)

Mode (P): ceftriazine (with 10 mug plate)

P(mm) S(mm)

18 3

16 3

15 4

49 10

X10 × 100 ═ 20.4% (considering 10 μ g of ceftriaxone)

49

49 10

X10 × 100 ═ 20.4% (considering 10 μ g of ceftriaxone)

49

No significant zone of inhibition was present, so the Minimum Inhibitory Concentration (MIC) was very close to 0.2. mu.L of LMSV-6.

Volume suppression zone applied to plate

5μL................................................42.0mm

1.6M................................................20.0mm

A

Results and discussion

Rats were infected intraoperatively, and then the infection was allowed to progress and successfully treated with Securolide. The high activity of these lactones against E.coli, Klebsiella pneumoniae, Pseudomonas aeroginosa, Staphylococcus aureus and their low molecular weight are advantageous. These tests clearly show that Securolide has a high activity against Pseudomonas, one of the microorganisms difficult to destroy.

Example 2: determination of antifungal Activity

Materials and methods:

female mice weighing 18 to 22g were used and an amount of Candida albicans 44858 was administered to the tail vein at a rate of 106CFU (CFU: colony forming unit) per mouse. Mice were divided into five batches for five batches and treated in the following manner:

1 hour after infection:

a first group: mice were dosed orally with the product at 25mg/kg,

second group: mice were dosed intraperitoneally with the product at 25mg/kg,

third group: mice were orally administered ketoconazole at 25mg/kg,

and a fourth group: mice were given ketoconazole intraperitoneally at a dose of 25mg/kg,

and a fifth group: the mice did not receive any antifungal treatment.

The number of dead mice was counted after 22 days.

Results and discussion

The activity of the product is very excellent in both modes of administration. The same treatment is also effective in the "epidermal mode" with dermatophytes, such as trichophyton, and in the sub-lethal mode.

Minimum Inhibitory Concentration (MIC)

Candida albicans cells were prepared as described in J.Antimicobiological chemistry, 38, 579-587, and washed 3 times with 0.1M phosphate solution and immediately used to determine the Minimum Inhibitory Concentration (MIC). MIC was determined by modifying microplates according to standard methods of the Comite National laboratory clinical standards.

RPMI-1640 and L-glutamic acid were treated with 0.15M MOPS (3- [ N-morpholino) pH7]Propanesulfonic acid) solution. Candida albicans cell (1.5X 10)³cells/mL) were added to the wells of a 96-well plate containing RPMI-1640 and a dilution of the antifungal agent. Results were read after 48 hours incubation at 35 ℃ and the MIC or minimum inhibitory concentration that inhibited Candida albicans cell growth was determined.

Minimum fungicidal concentration

After 48 hours the MIC is read, the plate is shaken and a 10. mu.L aliquot is pipetted from the well into a rectangular plate containing glucose. Plates were incubated at 35 ℃ for 48 hours and the lowest fungicidal and antifungal concentrations at which no colony forming units appeared were determined.

Example 3 cytotoxic or anti-tumor assays

The antiproliferative and cytotoxic effects of the drug can be quantified by using the tetrazolium salt, MTT (3- (4, 5-dimethylthiazol-2-yl) -2, 5-biphenyltetrazolium bromide). In this colorimetric assay, the yellow tetrazolium salt, MTT, is broken down into purple formazan by the mitochondrial enzyme, succinate dehydrogenase, in living cells. Formazan accumulates in living cells because it cannot permeate the cell membrane. The amount of formazan produced is proportional to the amount of viable cells after drug treatment. This assay was used to test our compounds for cytotoxic and anti-tumor effects on a variety of cancer cell lines.

Previous methods have employed well-known tissue culture techniques using a variety of cancer cell lines, such as HEP-2 (laryngeal carcinoma), HELA (cervical carcinoma). Formazan concentration was measured as a concentration gradient Securolide by a multi-well scanning spectrophotometer (ELISA scanner). Subsequent statistical data processing allowed the establishment of a semi-Inhibitory Concentration (IC) as a quantitative parameter of antitumor activity₅₀)。

The method and the material are as follows:

materials:

-culture medium (DMEN + all)

EDTA (Quantant reagent captures Ca present in cell membranes⁺⁺/Mg⁺⁺Ions, promoting detachment of cell membranes from the plate)

-trypsin-EDTA

-DMEM + ALL and 10% TERNERO RECENTAL serum.

Dimethyl sulfoxide (DMSO, inert solvent)

-MTT (3- (4, 5-dimethylthiazol-2-yl) -2, 5-biphenyltetrazolium bromide

Multi-well scanning spectrophotometer (ELISA scanner)

Anti-tumor assay

According to Skehan et al j.nat. cancer inst.82: 1107(1990) to determine an assessment of growth of suppressor cells. Cells were plated at 400 to 1200 cells per well in 96-well plates and incubated for 15-18 hours at 37 ℃ before adding drug to detach the cells from the plate. The compounds tested were dissolved in 100% DMSO and further diluted with RPMI-1640 containing 10mM HEPES. Each cell line was treated with 10 concentrations (5 log scale range) of compound. After 72 hours of incubation, 100mL of ice-cold 50% TCA was added to each well and incubated at 4 ℃ for 1 hour. The plates were then washed 5 times with tap water to remove TCA, low molecular weight metabolites and serum proteins. Sulforhodamine B (SRB) (0.4%, 50mL) was added to each well. After 5 minutes incubation at room temperature, the plates were washed 5 times with 0.1% acetic acid and air dried. Bound dye was dissolved with 10mM Tris base (pH10.5) for 5 minutes on a rotary shaker. The optical density was measured at 570 nm.

The data were analyzed using a Sigmoid-Emax concentration response model (see Holford, N.H.G.: Scheiner, LB., "Understanding dose-response relationships: pharmacokinetic-pharmacodynamic models", (Understanding the dose-response relationship: Clinical applications of pharmacological-pharmacological models), Clin. Pharimoackin 6: 429-Pharmacozac 453(1981) with nonlinear regression, weighted by the inverse square of the predicted response analysis software was developed by the Roell Swarkan research using Microsoft's FORTRAN language, employed by Nash (see NaNash. J.C., algebra., "computer compression n. mu. Merics method: linear and functionalMinimize "" Compact n μ Mericsal method for computers: linear algabra and function simulation, "John Wiley&Sons, New York, 1979) Marquardt algorithm for nonlinear regression (see Marquardt, d.w., "algorithm for nonlinear parameter least squares prediction" "An algorithm for least squares estimation of nonlinear parameters," j.soc.ind. "). Calculated to give 50% growth Inhibition (IC)₅₀) The drug concentration of (a).

Example 4 in vitro cytotoxicity of LMSV-6 against cervical and prostate cancer cells 4 tumor cell lines including human prostate cancer PC-4 (androgen insensitive) and LnCaP (androgen insensitive) and human cervical cancer CaSKi (human papillomavirus (HPV) type 16 positive and C-33 (HPV-negative) at 5% CO₂Cultured at 37 ℃ in RPMI-10% bovine serum, glutamic acid and antibiotics.

LMSV-6 vs C-33 (LD) was recorded after 4 hours₅₀538 μ M) were tested (at 1-1000 μ g/mL or 10.2-10, 200 μ M), but not the other (LD)₅₀> 1, 982. mu.M). After 72 hours the LMSV-6 concentration was 1X 10^-2To 10²μ M was shown to have antiproliferative activity against all cell types. The lethal doses for each cell line were as follows:

cell LD50(μ M)

C-33 7.10

LNCaP 26.5

PC-4 59.2

CaSKi 64.3

In general, methotrexate, doxorubicin and paclitaxel (LC)₅₀0.1 μ M) inhibits the proliferative activity of all types of cancer at concentrations below 0.001 to 0.5 μ M. The only exception was methotrexate, which was not active against CaSKi.

Tamoxifen was found to be cytotoxic to all four cell types (LC measured)₅₀34.5-79.7 μ M, compared to the expected 50 μ M). The curve shapes representing the percent inhibition of proliferation for 1000 μ g LMSV-6 and 1000 μ M tamoxifen are similar and parallel, with tamoxifen having an intensity of 1.53 times that of LMSV-6 (LD)₅₀345 μ M and 528 μ M, respectively).

Example 5 anti-helicobacter pylori Activity

Agar dilution of antimicrobial compounds

6mg of the compound to be evaluated was dissolved in 0.6mL of 100% dimethyl sulfoxide (DMSO) and then supplemented to 6mL with sterile Brucella medium and the solubility recorded. The final concentration of DMSO was 10% of the total volume. Serial 2-fold dilutions were made in sterile brucella broth (3mL Securolide +3mL brucella broth). 2mL aliquots of each medium dilution within the series were placed in separate sterile petri dishes and 18mL of thawed and cooled (approximately 50 ℃) Brucella agar supplemented with 7% horse blood was added. This resulted in a final 1: 10 dilution of Securolide in agar, and a final concentration of 1% DMSO. For example, the high difficulty of drugs in agar is 100. mu.g/mL. Agar plates were prepared the day before inoculation and stored in the refrigerator overnight.

Inoculation preparation

Helicobacter pylori was cultured on tryptic soy-5% sheep blood agar plates, and subcultured every 48 hours. Helicobacter mustelae (Helicobacter mustelae) was cultured on the same agar and passaged every 48-60 hours depending on the growth of the previous passage. Plates were grown at 37 ℃ in GasPak jars with water activated (10mL) CampyPak Plus (BBL microbial System) envelopes with palladium catalysts.

Helicobacter cultures were cultured in deep petri dishes in 10mL of brucella broth supplemented with 10% fetal bovine serum. Plates were incubated at 37 ℃ in GasPak jars with water activated (10mL) CampyPak Plus envelopes with palladium catalyst on a shaker at 50 rpm.

Overnight culture (approximately 10)⁸CFU/mL) was diluted 10-fold in brucella medium (no FCS) in a screw-capped tube for use as a standard inoculum. The wells of the Steer replicator were filled with 0.8mL of diluted organism and approximately 0.002mL volumes of 2X 10⁴The individual cells were placed on the agar surface. The inoculated plates were placed in GasPak jars with a palladium catalyst-loaded coating of Campy Pak Plus (BBL microbial System) Plus water activated (10mL) and incubated for 48 hours at 37 ℃.

Results and discussion

After the end of the culture, all test plates were compared to the growth control plate without Securolide. MIC is the concentration that inhibits growth compared to control plates. The thin grown film visible to the naked eye at higher concentrations was ignored and not considered as a true MIC. Control organisms were also inoculated on each plate and diluted 1000-fold for use as inoculum. Control organisms include selected cultures of Campylobacter jejuni (Campyylobacter jejuni), Escherichia coli [ ATCC 35218, Lote 202602, Exp 05/2000(19-258) ], Pseudomonas aerogenes [ ATCC 27853, Lote 202992, Exp 08/2000(19-060) ], Enterobacter cloacae (E.Cloacae), providencia stuartii (providedica) and providencia rapacipense (P.retti). The plate and/or inoculum passage should not be outside the microaerobic microbial environment for more than 2 hours. All operations involving helicobacter cultures were performed under a laminar flow hood to reduce the chance of mold contamination of the culture.

Lee et al, Gastroenterology, 99: 1315-23(1990), which are used for predicting the activity of an anti-helicobacter pylori compound in a human body. Helicobacter felis (helicobacter felis) was cultured in brucella broth containing 10% fetal bovine serum. The frozen cultures were thawed rapidly, the cultures were examined for motility, and 0.5cc of the thawed frozen culture was plated onto deep tissue culture dishes containing 9.5cc of brucella/serum mixture. The plates were placed in a Capy Pak tank [ BBL ] to ensure a microaerobic environment. The jar was placed on a rotary shaker at 60rpm and 37 ℃ in an incubator. After 18 hours, a visible haze should appear. The purity and motility of the culture were checked under a (phase) microscope and then collected into flasks. Swiss-Webster female mice (18-20g) were fasted for 18 hours prior to infection. Mice received a total of three infections every other day for a week. Dosing began 2 weeks after the last administration of the organism. Treatment was performed once daily for 14 consecutive days. Mice were sacrificed approximately 3 weeks after the end of treatment. For each mouse, its stomach was excised and opened along the greater curvature. A column (3mm tissue section) was taken from the antrum area. After the surface of the column was washed, it was cut up and dropped into a test tube containing 100. mu.l of urease reagent. The urease reagent (pH6.3-6.5) contains urea and phenol red. If helicobacter is present, urease will break down urea causing a pH change. Purple (alkaline) indicates positive helicobacter, and red/yellow (no change) indicates negative helicobacter. Any color change was recorded after 18 hours. Approximately 20 mice per treatment group, the percentage of positives in each group was recorded.

There are several methods used clinically to determine the presence of H.pylori in humans, and to determine the success of a treatment to eliminate the organism from the patient, prior to treatment, as a preliminary diagnosis of infection.

The urea breath test involves the uptake of radiolabeled urea. Helicobacter produces urease, which degrades urea, and is not contained by mammalian gastric cells. Thus exhalation of labelled carbon dioxide (mass spectrometry or radioactivity depending on the isotope used) indicates the presence of H.pylori.

Serology can also be used to assess helicobacter pylori infection. Antibodies to H.pylori, such as IgG and IgA, are detected in serum using enzyme-linked immunosorbent assay (ELISA). A variety of different H.pylori proteins can be used as antigens.

Endoscopy of patients provides tissue samples that can be cultured in microaerophilic environments to diagnose helicobacter pylori infection. Alternatively, the sample may optionally be examined histologically using one of a variety of dyes, such as giemsa or hematoxylin-eosin. Urea detection can also be used, which also takes advantage of the urease production by H.pylori. This detection results in an observable change in pH based on ammonia produced by urea hydrolysis.

Example 6 analysis of anti-gingivitis Activity

Evaluation of oral compositions against gingivitis, a study of 30 beagle dogs (beagle dog) using a Securolide-containing mouth rinse for a period of 10 weeks clearly shows its superior efficacy against gingivitis. The method used included complete removal of hard and soft dental deposits, followed by feeding the dogs on a soft diet for 6 weeks to allow gingivitis to develop. The dentition was then treated twice daily for five days a week with the test solution for approximately 15 seconds on each side of the oral cavity. Animals were examined and scored for the degree of inflammation of gingivitis according to a scale of 0 to 3:

0 is the inflammation-free condition, and 0 is the inflammation-free condition,

1, the slight local edema and redness of the gingival margin, no bleeding caused by light pressure of the fingers,

2, moderate edema and redness of the gingival margin, bleeding under light pressure of the fingers,

severe edema and ulceration of the gingival margin and attached gingiva also bleeds without light finger pressure. Placebo and 0.5% metronidazole washes were used as negative and positive controls, respectively.

Results and discussion

Compared to placebo, the Securolide lotion significantly reduced the development of gingivitis. The product can effectively control gingivitis and treat periodontal disease. Furthermore, a simple method of improving oral hygiene is provided, i.e. applying the product regularly to the oral cavity 1 to 3 times per day.

EXAMPLE 7 in vitro mutagenesis study (Ames test) of LSV-6

The study was conducted according to the standard of USC 79/831, which established a guideline for a toxic substance analysis method.

The Ames test detects drug-induced back mutations in mutant Salmonella typhimurium strains that carry the rfa mutation to render their cell membranes permeable to chemical agents. Each test line carries a known mutation (deletion, substitution or addition of base pairs) in the histidine operon. The presence of these mutations rendered these cells unable to grow in histidine-deficient medium. Drug-induced reversion mutations reverse the effects of all these mutations, restoring the strain to its previous histidine-positive state, and thus, enhancing Salmonella typhimurium growth on histidine-deficient media.

A drug is considered mutagenic if it increases the number of colonies more than two-fold on histidine-deficient medium. Addition of LMSV-6(1 or 10. mu.g/mL) to bacterial strain TA98 did not increase the number of normal colonies on histidine-deficient medium, while addition of LMSV-6 (< 1000. mu.g/mL) to bacterial strain TA100 did not increase the number of colonies, and therefore met the criteria for mutagenesis. These preliminary results indicate that LMSV-6 does not reverse mutate Salmonella typhimurium TA98 or TA100 and cannot be expected to cause mutations in the human genome.

Example 8 study of the adverse Effect of LMSV-6 on healthy volunteers

Method

19 healthy volunteers participated in the study. The age of the participants was between 21 and 40 years, with a mean age of 30.5 ± 7.2 years. Weight was between 150 and 190 pounds, median 163.2 + -14.3 pounds, height was between 1.8 and 2.1 meters, and average 1.9 + -0.1 meters. Subjects were randomly selected according to national and international standards and established human study specifications.

The health status of the participants was established using several clinical assessments, laboratory tests, electrocardiogram and chest X-ray. The function of the liver and kidney system was examined by chemoenzymatic studies. Also, blood chemistry tests, blood pathology tests and urinalysis were performed on each participant. For females, no pregnancy and/or no lactation was determined by laboratory tests and obstetrics evaluation.

Random selection divided participants into 3 groups (n ═ 6):

the first group (placebo) received 2.0mL of physiological saline (CINa)⁺，0.9％)，

The second group, received an intramuscular injection of 60mg of LMSV-6, and

the third group (n ═ 7) received an intramuscular injection of 100mg of LMSV-6.

The participants received several tests that were used to establish the base value of the objective parameters of the analysis, including:

laboratory examination: hemogram, hematochemistry, urinalysis and stool analysis

Cardiovascular function: arterial Tone (TA), heart Rate (RC), heart rate (FC), and radial artery beat (PR)

Lung function: lung respiration (VP) and respiratory rate (FR)

Renal function: volume and Frequency of Urine (FU)

Sensory system: auditory, visual, olfactory, gustatory and sensory reflexes

Skin and/or integuments: sensitivity, skin texture, temperature and musculoskeletal flexibility

Autonomic nervous system function: salivary and sweat gland activity, gastrointestinal mortality, and visceral reflex hypersensitivity: local sensitivity and systemic sensitivity.

The evaluation was performed at the following time intervals: times 0.0, 5.0, 15.0, 20.0, 30.0, and 45.0 minutes; 1.0 hour, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0, 12.0, 24.0, 36.0 and 48.0 hours.

Results

Laboratory tests and special examinations to verify the health status of the participants showed that these values remained at the baseline level and that no inconsistency with the placebo group was observed.

Laboratory tests of urine function and microbiological patterns found values within the reference values of the method used. Similarly, the results of stool analysis also showed no pathological changes.

Individual values and mean standard deviations of the relative cardiovascular parameters including arterial Tension (TA), heart rate (FC), radial Pulsation (PR) and heart Rhythm (RC) were measured for each group of participants before and during the experiment to determine side effects and tolerance limits for LMSV-6. For healthy volunteers, they received 60 and 100mg of LMSV-6 and 2mL of 0.9% sodium chloride as placebo, and the results showed that the levels of TA, FC, PR and RC did not change with treatment compared to the baseline values and those of the placebo control group. Similarly, there was no change in the electrocardiogram under the basal conditions and after the study.

Individual values and standard mean deviations for each group with respect to respiratory function were also measured for evaluating the effect of LMSV on healthy volunteers. The results show that the respiratory function and kinetics of the participants were not altered after treatment with the experimental dose compared to the basal value and placebo control group. The respiratory rate (FR) was maintained throughout the test in the range of 18 to 20 breaths per minute.

According to observations under basal conditions and in the placebo control group, the parameters evaluated with respect to respiratory kinetics, inspiration, expiration, tracheal respiration, bronchial respiration and chest-lung expansion and upper and lower airflow were not altered.

The evaluation of the effects of LMSV-6 with reference to renal function parameters showed that the urinary frequency values remained within the range of 3.1 ± 0.7-fold to 3.3 ± 1.5-fold without significant difference observed directly within 12 hours in the individuals of the control group and the individuals of the treatment group. Similarly, the mean urine volume was 434.2. + -. 213.2, 489.2. + -. 94.3 and 394.3. + -. 103.9mL in the placebo control group and the group receiving 60 and 100mg of LMSV-6, respectively, over the same period of time. This indicates that the treatment did not affect basic renal function.

Data on the effect of LMSV-6 on mental state and sensory accuracy: auditory, visual, olfactory and gustatory discriminatory power indicates that treatment does not cause a change in any of the functional or analytical parameters described above. Similarly, the superficial and tendon reflexes do not change anything.

The data for the evaluation of the effects of LMSV-6 on skin sensitivity and temperature show that the treatment did not cause any change in the skin sensitivity or temperature of the participants. Similarly, there was no evidence of any change in musculoskeletal contractility in any of the participants in the control and treatment groups.

The treatment did not produce any change in the texture and moisturization of the skin of all participants in this study, nor did it produce changes in the properties of the oral and nasal mucosa as a result of the treatment.

The relevant data for the assessment of local and systemic anaphylaxis (reactivity) indicate that no anaphylaxis was shown in any of the participants.

Data from the evaluation of the effects of LMSV-6 on the level of autonomic nerves (glands and viscera) showed no change in autonomic nerves at the gastrointestinal visceral level, the level of genito-urinary, the larynx-eye glands, or cardiotonic effect over the range of study doses (60 and 100 mg); that is, there is no change at the exocrine and endocrine gland levels.

The results of this study confirmed that LMSV-6 had sufficient tolerability limits, and that all participants were tolerated at the maximum dose tested (100 mg). Similarly, there was no evidence, sign or symptom of adverse effects in all participants who participated in the study.

Claims (31)

1. A process for preparing a compound of formula Ia, Ib or Ic comprising:

providing a precursor having a lactone structure, and

the precursor is reacted with one or more chemical reagents to provide a product having one of the general formulas Ia, Ib, and Ic.

2. The compound of claim 1 wherein the precursor has the structure of formula II:

formula II

Wherein R is₁And R₂Independently a hydrogen atom or a group selected from: alkyl, substituted alkyl, allyl, substituted allyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halogen, hydroxy, alkoxy, substituted alkoxy, alloxy, phenoxy, substituted phenoxy, aryloxy, substituted aryloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C₁-C₂₀Cyclic, substituted C₁-C₂₀Rings, heterocycles, substituted heterocycles, amino acids, peptides and polypeptide groups.

3. An isolated or synthetic compound defined by one of the following structures:

formula Ia formula Ic

Wherein R is₁-R₆Independently a hydrogen atom or a group selected from: alkyl, substituted alkyl, allyl, substituted allyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,Phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, alkoxy, substituted alkoxy, alloxy, phenoxy, substituted phenoxy, aryloxy, substituted aryloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C₁-C₂₀Cyclic, substituted C₁-C₂₀Rings, heterocycles, substituted heterocycles, amino acids, peptides and polypeptide groups;

z is a heteroatom selected from the group consisting of oxygen, sulfur or nitrogen radicals in a linear, branched or cyclic form; and

x is a heteroatom selected from the group consisting of oxygen, sulfur or nitrogen radicals in a linear, branched or cyclic form.

4. A compound according to claim 3 selected from compounds 1 to 50 as defined in table 1.

5. A pharmaceutical composition comprising a lactone compound or a pharmaceutically acceptable salt or hydrate thereof and a physiologically acceptable carrier, wherein the lactone compound has one of the following structures:

formula Ia formula Ic

Wherein R is₁-R₆Independently a hydrogen atom or a group selected from: alkyl, substituted alkyl, allyl, substituted allyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halogen, hydroxy, alkoxy, substituted alkoxy, alloxy, phenoxy, substituted phenoxy, aryloxy, substituted aryloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C₁-C₂₀Cyclic, substituted C₁-C₂₀Rings, heterocycles, substituted heterocycles, amino acids, peptides and polypeptide groups;

z is a heteroatom selected from the group consisting of oxygen, sulfur or nitrogen radicals in a linear, branched or cyclic form; and

x is a heteroatom selected from the group consisting of oxygen, sulfur or nitrogen radicals in a linear, branched or cyclic form.

6. The pharmaceutical composition of claim 5, wherein the lactone compound is selected from the group consisting of compounds 1-50 as defined in Table 1.

7. The pharmaceutical composition of claim 5, wherein the lactone compound is effective as an antibacterial agent, an antifungal agent, an antineoplastic agent, an anti-inflammatory agent, an anti-gingivitis agent, or an anti-periodontitis agent.

8. The pharmaceutical composition of claim 6, wherein the lactone compound is effective as an antibacterial, antifungal, antitumor, anti-inflammatory, or anti-periodontitis agent.

9. The pharmaceutical composition of claim 7, wherein the lactone compound is effective as an anti-inflammatory agent for treating or preventing an inflammatory disease selected from the group consisting of: arthrosclerosis, pulmonary fibrosis, systemic lupus erythematosus, pancreatitis, sarcoidosis, glomerulonephritis, and organ transplant rejection.

10. The pharmaceutical composition of claim 8, wherein the lactone compound is effective as an anti-inflammatory agent for treating or preventing an inflammatory disease selected from the group consisting of: arthrosclerosis, pulmonary fibrosis, systemic lupus erythematosus, pancreatitis, sarcoidosis, glomerulonephritis, and organ transplant rejection.

11. The pharmaceutical composition of claim 7 wherein the lactone compound is effective as an antineoplastic agent for treating or preventing cancer.

12. The pharmaceutical composition of claim 8, wherein the lactone compound is effective as an antineoplastic agent for treating or preventing cancer.

13. The pharmaceutical composition of claim 7, wherein the lactone compound is effective as an antitumor agent for treating or preventing a cancer selected from the group consisting of: melanoma, leukemia, breast cancer, lung cancer, ovarian cancer, colon cancer, esophageal cancer, liver cancer, and lymphoma.

14. The pharmaceutical composition of claim 8, wherein the lactone compound is effective as an antitumor agent for treating or preventing a cancer selected from the group consisting of: melanoma, leukemia, breast cancer, lung cancer, ovarian cancer, colon cancer, esophageal cancer, liver cancer, and lymphoma.

15. The pharmaceutical composition of claim 7, wherein the lactone compound is an antibacterial agent effective against Helicobacter pylori (Helicobacter pylori) for treating or preventing gastric ulcer, gastritis, dyspepsia or gastric cancer.

16. The pharmaceutical composition of claim 8, wherein the lactone compound is an antibacterial agent effective against helicobacter pylori, for treating or preventing gastric ulcer, gastritis, dyspepsia or gastric cancer.

17. The pharmaceutical composition of claim 7, wherein the lactone compound is effective against gingivitis and/or periodontitis, wherein the composition is an oral composition further comprising another anti-gingivitis agent effective against the gram-negative anaerobic microorganisms Bacteriodes assaccharolyticus, Bacteriodes gingivalis, and mixtures thereof.

18. The pharmaceutical composition of claim 8, wherein the lactone compound is effective against gingivitis and/or periodontitis, wherein the composition is an oral composition further comprising another anti-gingivitis agent effective against the gram-negative anaerobic microorganisms Bacteriodes assaccharolyticus, Bacteriodes gingivalis, and mixtures thereof.

19. The pharmaceutical composition of claim 9, wherein the organ transplant rejection is a rejection of an organ transplant selected from the group consisting of: kidney, liver, lung and heart transplantation.

20. The pharmaceutical composition of claim 10, wherein the organ transplant rejection is a rejection of an organ transplant selected from the group consisting of: kidney, liver, lung and heart transplantation.

21. The pharmaceutical composition of claim 7, wherein the compound is effective against digestive, urinary, reproductive or cutaneous candidiasis, cryptococcosis, bronchopulmonary or pulmonary aspergillosis, or invasive aspergillosis of the immunosuppressive system.

22. The pharmaceutical composition of claim 8 wherein the compound is effective against digestive, urinary, reproductive or cutaneous candidiasis, cryptococcosis, bronchopulmonary or pulmonary aspergillosis, or invasive aspergillosis of the immunosuppressive system.

23. A method for treating or preventing a disorder comprising administering to an animal a composition comprising an effective amount of a compound or a pharmaceutically acceptable salt or hydrate thereof, wherein the compound has one of the following structures:

formula Ia formula Ic

Wherein R is₁-R₆Independently a hydrogen atom or a group selected from: alkyl, substituted alkyl, allyl, substituted allyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halogen, hydroxy, alkoxy, substituted alkoxy, alloxy, phenoxy, substituted phenoxy, aryloxy, substituted aryloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C₁-C₂₀Cyclic, substituted C₁-C₂₀Rings, heterocycles, substituted heterocycles, amino acids, peptides and polypeptide groups;

z is a heteroatom selected from the group consisting of oxygen, sulfur or nitrogen radicals in a linear, branched or cyclic form; and

x is a heteroatom selected from the group consisting of oxygen, sulfur or nitrogen radicals in a linear, branched or cyclic form;

wherein the disorder is selected from: cancer, gingivitis, periodontitis, bacterial diseases, fungal diseases, and inflammatory diseases.

24. The method of claim 23, wherein the compound is selected from the group consisting of compounds 1-50 as defined in table 1.

25. The method of claim 23, wherein the cancer is selected from the group consisting of melanoma, leukemia, breast cancer, lung cancer, ovarian cancer, colon cancer, esophageal cancer, liver cancer, and lymphatic cancer.

26. The method of claim 23, wherein the bacterial disease caused by helicobacter pylori is selected from the group consisting of gastric ulcer, gastritis, dyspepsia and gastric cancer.

27. The method of claim 23, wherein the inflammatory disease is selected from the group consisting of: arthrosclerosis, pulmonary fibrosis, systemic lupus erythematosus, pancreatitis, sarcoidosis, glomerulonephritis, and organ transplant rejection.

28. The method of claim 23, wherein the organ transplant rejection is a rejection of an organ transplant selected from the group consisting of: kidney, liver, lung and heart transplantation.

29. The method of claim 23 wherein the disorder is gingivitis or periodontitis, and

wherein the composition is a topical formulation for daily application to the oral cavity.

30. The method of claim 23, wherein the condition is gingivitis or periodontitis, and

wherein the composition is a topical formulation for daily application to the oral cavity.

31. The method of claim 23, wherein the fungal disease is selected from the group consisting of: candidiasis of digestive organs, urinary organs, reproductive organs and skin, cryptococcosis, bronchopulmonary and pulmonary aspergillosis, and invasive aspergillosis of the immunosuppressive system.