Title of Invention


Abstract A composition for the treatment or prevention of a condition selected from the group consisting of pain, an ocular condition, a neurodegenerative condition, and a sympathetically-enhanced stress-associated condition, comprising: a) 0.001 to 5% w/w of a first component comprising a compound whose activity results in a direct or indirect activation of the alpha 2 adrenergic receptor, and b) 0.001 to 5% w/w of a second component comprising an alpha 1 adrenergic receptor antagonist, in apharmaceutically acceptable carrier.
This invention relates to compositions and methods for treating malaria. Specifically, this invention relates to pharmaceutical compositions including spiro and dispiro trioxolanes, and methods of their use and manufacture
Malana is an acute and often chronic infectious disease resulting from the presence of protozoan parasites within red blood cells Caused by single-celled parasites of the genus Plasmodium, malana is transmitted from person to person by the bite of female mosquitos
Although once prevalent in North Amenca and other temperate regions of the world, today malaria occurs mostly in tropical and subtropic countnes. Each year, between 400 million and 600 million people contract the disease, and 1 5 million to 2 7 million die of the disease
Four species of Plasmodium protozoan parasites are generally responsible for malana, including Plasmodium vivax, Plasmodium falciparum, Plasmodium malariae, and Plasmodium ovale Of the four, Plasmodium falciparum is the most dangerous, accounting for half of all clinical cases of malana and 90% of deaths from the disease
The transmission of malana begins when a female mosquito bites a human already infected with the malana parasite. When the infected mosquito bites another human, sporozoites in the mosquito's saliva are transferred into the blood, which then travel to the liver In the liver, the sporozoites divide rapidly, then enter the bloodstream where they invade red blood cells Inside these blood cells, the merozoites multiply rapidly until they cause the red blood cells to burst, releasing into the blood stream a new generation of merozoites that then infect other red blood cells
The symptoms associated with malana are generally associated with the bursting of the red blood cells The destruction of the red blood cells spills wastes, toxin, and other debns into the blood This in turn causes an intense fever that can leave the infected individual exhausted and bedndden More severe symptoms associated with repeat infections and/or infection by Plasmodium falciparum include anemia, severe headaches, convulsions, delinum and, in some instances, death
The treatment of malaria has been especially difficult due to the ability of malaria parasites to develop resistance to drugs. Quinine, an antimalanal compound that is extracted from the bark of the South American cinchona tree, is one of the oldest and most effective pharmaceuticals in existence The downside to quimne is that it is short-acting, and fails to prevent disease relapses. Further, quinine is associated with side effects ranging from dizziness to deafness
Chloroquine is a synthetic chemical similar to quinine It became the drug of choice for malaria when it was developed in the 1940s due to its effectiveness, ease of manufacture, and general lack of side effects However, in the last few decades, malaria parasites in many areas of the world have become resistant to chloroquine
Mefloquine is another synthetic analog of quimne that has been used in the treatment of malaria Malaria parasites have also developed resistance to mefloquine, however Mefloquine is also associated with undesirable central nervous side effects in some patients, including hallucinations and vivid nightmares
Antifolate drugs are effective against malaria parasites by inhibiting their reproduction Although the parasites have also developed a resistance to antifolate drugs, the drugs can still be used effectively in combination with other types of antimalanals The use of combination therapies in treating malaria has the drawbacks of being inconvenient and expensive, however
More recent developments in the treatment of malana have involved the use of the peroxide functional group, as exemplified by the drug artemisinm, which contains a unique 1,2,4-tnoxane heterocyclic pharmacophore. The antimalanal action of artemisinm is due to its reaction with the iron in free heme molecules in the malana parasite with the generation of free radicals leading to cellular destruction.
The discovery of artemismin (qinghaosu), a naturally occurring endoperoxide sesquiterpene lactone (Meshnick et al, 1996; Vroman et al 1999, Dhingra et al, 2000) initiated a substantial effort to elucidate its molecular mechanism of action (Jefford, 1997, dimming et al, 1997) and to identify novel antimalanal peroxides (Dong and Vennerstrom, 2001) Many synthetic 1,2,4-tnoxanes, 1,2,4,5-tetraoxanes, and other endoperoxides have been prepared
Although the clinically useful semisynthetic artemisinin derivatives are rapid acting and potent antimalarial drugs, they have several disadvantages including recrudescence, neurotoxicity, (Wesche et al, 1994) and metabolic instability (White, 1994) A fair number of these compounds are quite active in vitro, but most suffer from low oral activity (White, 1994; van Agtmael et al, 1999). Although many synthetic antimalarial 1,2,4-tnoxanes have since been prepared (Cumming et al., 1996, Jefford, 1997), there exists a need in the art to identify new peroxide antimalarial agents, especially those which are easily synthesized, are devoid of neurotoxicity, and which possess improved pharmacokinetic properties, e g improved stability, oral absorption, etc
Accordingly, it is a primary objective of the present invention to provide compositions and methods for prophylaxis and treatment of malana using spiro and dispiro 1,2,4-tnoxolanes
It is a further objective of the present invention to provide a composition and method for prophylaxis and treatment of malana using spiro and dispiro 1,2,4-tnoxolanes that is nontoxic
It is a further objective of the present invention to provide a composition and method for prophylaxis and treatment of malana using spiro and dispiro 1,2,4-tnoxolanes that is metabohcally stable and orally active.
It is yet a further objective of the present invention to provide a composition and method for prophylaxis and cost-effective treatment of malana using spiro and dispiro 1,2,4-tnoxolanes that do not involve a treatment regimen of more than three days
It is a further objective of the present invention to provide compositions and methods for prophylaxis and treatment of malana using spiro and dispiro 1,2,4-tnoxolanes that can be used either as stand-alone medicaments or in combination with other agents
It is still a further objective of the present invention to provide novel intermediates for synthesizing compositions for prophylaxis and treatment of malana
The method and means of accomplishing each of the above objectives as well as others will become apparent from the detailed descnption of the invention which follows hereafter
The invention describes a method and composition for treating malaria with spiro and dispiro 1,2,4-tnoxolanes, their prodrugs and analogues. The tnoxolanes of this invention are stencally hindered on one side of the trioxolane heterocycle in order to provide chemical and metabolic stability to the trioxolane nng for better in vivo activity In one embodiment, the spiro and dispiro tnoxolanes are sterically hindered with an unsubstituted, mono-, di-, or poly-substituted C5-C12 spiro cycloalkyl group, which may be spiroadamantane In this embodiment, the spiro and dispiro tnoxolanes may include a spirocyclohexyl that is functionalized or substituted at the 4-position or a spiropipendyl nng that is functionalized or substituted at the nitrogen atom. In another embodiment, the tnoxolanes of this invention include an alkyl bndge from the 4-position of the spirocyclohexyl nng connecting a substituent that is most preferably a weak base The invention embraces achiral, achiral diastereomers, racemic mixtures, as well as enantiomenc forms of the compounds
The tnoxolanes of this invention possess excellent potency and efficacy against Plasmodium parasites, and a low degree of neurotoxicity In addition, several of the tnoxolanes are suitable for both oral and non-oral administration. Moreover, in companson to artemisinin semisynthetic denvatives, the compounds of this invention are structurally simple, easy and inexpensive to synthesize, and can be used effectively alone or in conjunction with other antimalanals.
The present invention relates to the development of spiro and dispiro 1,2,4-tnoxolanes for use in the prophylaxis and treatment of malana. The present invention is predicated upon the unexpected discovery that tnoxolanes that are relatively stencally hindered on at least one side of the tnoxolane heterocycle provide metabolic and chemical stability to the tnoxolane nng, thereby providing better in vivo activity, especially with respect to oral administration
As used herein the term "prophylaxis-effective amount" refers to a concentration of compound of this invention that is effective in inhibiting or preventing infection and subsequent disease by malanal parasites. Likewise, the term "treatment-effective amount" refers to a concentration of compound that is effective m treating malana in terms of preventing an increase in the concentration of malanal parasites, decreasing the
ILconcentration of malanal parasites, and/or "cunng" a malana infection, 1 e survival for 30 days post-infection.
Tetrasubstituted trioxolanes are relatively stable peroxidic compounds based on literature precedent (Gnesbaum et al, 1997a; 1997b). This may be due, in part, to the lack of α-hydrogen atoms The present inventors have synthesized new compounds in the tnoxolane class having both supenor antimalanal potency and oral efficacy Furthermore, the compounds of this invention have low toxicity, and half-lives conducive to treatment of malana which are believed will permit short-term treatment regimens companng favorably to other artemismm-like drugs. These compounds may also be used in malaria prophylaxis
In previous application, the present inventors disclosed certain novel tetrasubstituted tnoxolanes having the following structural formula
(Formula Removed)
wherein R1, R2, R3, and R4 represent combinations of nng systems, acyclic systems, and functional groups that provide sufficient stenc hindrance about the tnoxolane nng in order to give the nng chemical and metabolic stability R1, R2, R3 and R4 may be the same or different, and may be a linear or branched alkyl, aryl, or alkaryl group which is optionally substituted In the alternative, R1 and R2 taken together and/or R3 and R4 taken together may form an ahcychc group which is optionally interrupted by one or more oxygen, sulfur or nitrogen atoms and which group is optionally substituted In no event may any of R1, R2, R3 or R4 be hydrogen
In one embodiment, the compounds include those whereby R1 and R2 taken together and/or R3 and R4 taken together is a mono- or di-substituted C5-C12 spirocycloalkyl group which is optionally interrupted by one or more oxygen, sulfur, or nitrogen atoms, and which group is optionally substituted. In another embodiment, R1 and R2 taken together or R3 and R4 is spiroadamantane
The present invention discloses a new embodiment of tnoxolane compounds having the following structure
(Formula Removed)
The spirocyclohexyl ring may be optionally interrupted by one or more oxygen, sulfur or nitrogen atoms In this regard, R1 and R2 may be the same or different, and may be hydrogen, substituted or unsubstituted linear or branched alkyl, aryl, and alkaryl groups and substituted or unsubstituted alicyclic groups that may be interrupted by one or more oxygen, sulfur or nitrogen atoms, substituted or unsubstituted aromatic or heterocyclic groups that may be interrupted by one or more oxygen, sulfur or nitrogen atoms, a hydroxy group, or a halogen. In one embodiment, R1 or R2 is an amide. It has been unexpectedly found that amide-containing substituents at the 4-position provide antimalarial compounds with good oral absorption, good antimalarial activity, and good pharmacokinetics, 1 e rates of absorption, metabolism, and elimination that are suitable and advantageous for the prophylaxis and treatment of malaria.
In another embodiment, the compounds of this invention have the following structural formula
(Formula Removed)
whereby R3 is (CH2)n-Y In this formula, Y represents a functional group that, in one embodiment, is non-acidic, and in another embodiment is a weak base The Y functional group may be an alkyl, ketone, acid, alcohol, amine, amide, sulfonamide, guamdine, ether,
lister, oxime, urea, oxime ether, sulfone, lactone, carbamate, semicarbazone, phenyl, or heterocycle In one embodiment, n = 1 The alkyl "bridge" group has been found to improve the metabohcally stability (i e. oral activity and pharmacokinetics) of the antimalarial compounds of this invention.
In another embodiment of the invention, the tnoxolanes are weak bases, which provide an ideal combination of high intrinsic potency and good oral activity Two promising tnoxolane structural subtypes are weak base amides of tnoxolane amine OZ209 and trioxolane acid OZ78 These compounds have one of the following two structural formulas:
(Formula Removed)
Other substituents at the 4-position of the spirocyclohexyl ring are also possible that fall within the scope of this invention The spirocyclohexyl nng may also be substituted at other positions besides the 4-position For instance, the inventors have synthesized several compounds substituted at the 2-position of the spirocyclohexyl nng that provide excellent antimalarial potency.
In another embodiment of this invention, the compounds include an alkyl group connecting the substituent at the 4-position to the spirocyclohexyl ring In one embodiment, the alkyl group is methyl or ethyl In another embodiment, the alkyl group is methyl The substituent may also be directly attached to the 4-position of the spirocyclohexyl ring
The present inventors have identified two orally active lead dispiro-1,2,4-tnoxolanes, OZ03 and OZ05
(Formula Removed)
These trioxolanes have IC50s between 1 and 5 ng/ml against P falciparum in vitro, and presumably possess good therapeutic indices as no toxicity is evidence for either compound in a neuroblastoma cell line or at single 640 mg/kg doses in mice in the Rane test These results contrast with published data (de Almeida Barbosa et al, 1992; 1996) disclosing the weak in vitro antimalarial potency of several tricyclic trioxolanes, the best of which has an IC50 of 2000 ng/ml against P falciparum in vitro
A notable feature of these trioxolanes in comparison to the artemisinin semisynthetic derivatives is their structural simplicity A potential advantage of trioxolanes over both tnoxanes (Jefford, 1997, Cumming et al., 1997) and tetraoxanes (Vennerstrom et al., 2000) is a more convement access to structurally diverse, non-symmetrical, and in many cases, achiral compounds
Below are several dispiro 1,2,4-trioxolanes synthesized in accordance with the teachings of this invention. "OZ" is an internal designation for these compounds that will be used throughout the remainder of the application for convenience The structures of OZ01-OZ270 have been previously disclosed in prior applications U S Serial No. 09/886,666 (U S Patent No 6,486,199) and PCT/02/19767, and are therefore not repeated here

(Structure Removed)

OZ Series 32 (OZ280-OZ288)
(Structure Removed)
OZ Series 33 (OZ289-OZ297)
(Structure Removed)
OZ Series 34 (OZ298-OZ306)

(Structure Removed)
OZ Series 35 (OZ307-OZ315)

(Structure Removed)
OZ Series 36 (OZ316-OZ324)
(Structure Removed)
OZ Series 37 (OZ325-OZ333)
(Structure Removed)

OZ Series 38 (OZ334-OZ342)
(Structure Removed)

OZ Series 39 (OZ343-OZ351)
(Structure Removed)
OZ Series 40 (OZ352-OZ360)
(Structure Removed)
OZ Series 41 (OZ361-OZ369)
(Structure Removed)
The prototype tnoxolanes of this invention are OZ03 and OZ05 Preferred compounds identified thus far include OZ03, OZ05, OZ11, OZ25, OZ27, OZ61, OZ71, OZ78, OZ127, OZ145, OZ156, OZ163, OZ175, OZ177, OZ179, OZ181, OZ189, OZ205, OZ207, OZ209, OZ210, OZ219, OZ227, OZ229, OZ235, OZ255, OZ256, OZ257, OZ263,
OZ264, OZ265, OZ266, OZ267, OZ268, OZ269, OZ270, OZ271, OZ277, OZ281, OZ279, OZ288, OZ289, OZ290, OZ296, OZ297, OZ298, OZ301, OZ305, OZ309, OZ315, OZ317, OZ319, OZ320, OZ323, OZ329, OZ333, OZ335, OZ336, OZ337, OZ338, OZ339, OZ343, OZ349, OZ351, OZ353, OZ354, OZ357, OZ358, OZ359, OZ365, and OZ368 The most preferred compounds are OZ78, OZ163, OZ181, OZ207, OZ209, OZ255, OZ256, OZ257, OZ263, OZ264, OZ267, OZ271,OZ277, OZ279, OZ301, OZ305, OZ315, OZ317, OZ319, OZ323, OZ329, OZ338, OZ339, OZ349, OZ351, OZ354, OZ357, OZ359, and OZ368, with OZ277 and OZ279 being the best of those compounds identified thus far In general, the highest in vitro potency against malarial parasites is obtamed for tnoxolanes functionahzed or substituted at the 4-position of the spirocyclohexyl ring As a general rule, non-symmetrical, achiral trioxolanes are also preferred
Notable features of these spiro and dispiro 1,2,4-tnoxolanes in comparison to the artemisimn semisynthetic derivatives are their structural simplicity and ease of synthesis For example, dispiro tnoxolanes may be easily synthesized by the coozonolysis of the O-methyl oximes of cycloalkanones in the presence of the requisite cycloalkanone denvatives according to the method of Gnesbaum et al (1997a, 1997b) as illustrated below for the symmetrical dispiro cyclohexyl tnoxolane

(Formula Removed)

If yields are low in this coozono lysis reaction, yields can improve dramatically when the O-methyloxime and ketone are "reversed " This novel procedure provides a uniquely convenient method to synthesize spiro and dispiro tnoxolanes Advantages of the oxime ether route over the alkene approach include convenient synthesis of starting matenals (oxime ethers vs tetrasubstituted alkenes), higher yield and selectivity of formation of desired tnoxolanes by the judicious selection of paired reaction substrates The tnoxolanes may be punfied by crystallization or by flash column chromatography Their structures and punty may be confirmed by analytical HPLC, 1H and 13C NMR, IR, melting point and elemental analysis
Formation of a trioxolane from an oxime ether and a ketone is presumed to be a three-step process. The sequence begins by the electrophilic addition of ozone to the oxime double bond to form a primary ozonide. Second, the very unstable primary adduct fragments to a reactive carbonyl oxide driven in part by the concomitant expulsion of the relatively stable methyl nitrite Third, the carbonyl oxide undergoes a [3 + 2] cycloaddition with a ketone to give the secondary ozonide or 1,2,4-tnoxolane. It remains to be
(Formula Removed)
determined whether this is a stepwise or a concerted recombination process
As illustrated above by the synthesis of OZ03, most of the new dispiro tnoxolanes contain a spiroadamantane and can be synthesized by the coozonolysis of adamantanone O-methyl oxime in the presence of the requisite cycloalkanone derivative The preferred reaction solvents for the coozonolysis reactions are hydrocarbon solvents such as pentane or cyclohexane, more polar solvents tend to decrease the yield of the reaction When ketones are not readily soluble in pentane or cyclohexane, a mixed solvent (pentane/methylene chloride) or methylene chloride alone may be used Several factors govern the ratio of oxime ether to ketone In some reactions, in order to avoid diperoxide (1,2,4,5-tetraoxane) formation, to preclude diozonide formation from diketones, and to promote the reaction with readily pentane soluble ketones, excess ketone (2 1) is used Most commonly in the discovery synthesis stage, and especially in cases where ketones are not readily soluble m pentane, expensive, or difficult to remove in the reaction workup, a 1 1 ratio of ketone to oxime ether may be used In large scale trioxolane syntheses, a 1 5-fold excess of oxime ether can be used to achieve higher conversions of ketones into the desired product tnoxolanes without causing purification problems
There are several examples of where post-ozonolysis transformations were used to obtain trioxolane target compounds difficult, or in some cases, impossible to obtain directly (Kashima et al, 1987) by the coozonolysis method Trioxolane tertiary alcohols OZ90 and
(Formula Removed)
OZ108 can be obtained by methyllithium treatment of tnoxolane ketone OZ05 and trioxolane ester OZ70, respectively. In other reactions, tnoxolane lactone OZ17 and tnoxolane alcohol OZ32 were obtained by treatment of OZ05 with m-CPBA and sodium borohydnde, respectively. In addition, vanous oxime ethers, hydrazones, ketals, and amines (reductive amination with sodium tnacetoxyborohydnde) were also obtained from tnoxolane ketone OZ05 in good to excellent yields In the examples noted above, it is evident that troxolane ketone OZ05 is a key intermediate as its ketone functional group provides a convenient means for functional group transformation
(Formula Removed)
Further evidence of the stability of these tnoxolanes to reducing agents is shown by the reduction of tnoxolane esters OZ70 and OZ61 mto then corresponding tnoxolane alcohols OZ119 and OZ89 with a mixture of lithium borohydnde and lithium tnethylborohydnde, and the hydrazmolysis of the tnoxolane phthahmides OZ136 and
(Formula Removed)
OZ146 into their corresponding tnoxolane amines OZ137 and OZ209
As shown in the examples below, tnoxolane esters can be conveniently converted into their corresponding tnoxolane acids
(Formula Removed)
In addition to tnoxolane ketone OZ05, tnoxolane amine mesylate OZ209, tnoxolane ester OZ61 and tnoxolane acid OZ78, tnoxolane alcohols OZ119 and OZ89 have and will continue to be key intermediates for post-ozonolysis synthetic transformations A recent example is the synthesis of tnoxolane triazole OZ177 in a reaction between the mesylate derivative of OZ119 and the sodium salt of 1,2,4-tnazole
(Formula Removed)
It has been found that the coozonolysis method using oxime methyl ethers offers a rapid, flexible, and predictable access to structurally diverse tnoxolanes In fact, several key tnoxolanes that have served as important building blocks have been prepared in large scale including OZ05 (100 mmol), OZ61 (100 mmol), and OZ146 (60 mmol), with no decrease in reaction yields over the usual 5-10 mmol scale. Furthermore, both OZ61 and OZ146 can be conveniently isolated as white solids by addition of ethanol to the crude reaction mixtures
Differential scanning calonmetry (DSC) experiments (Cammenga, and Epple, 1995) reveal that these compounds have good thermal stability, comparable to artemisinin The average Tm, dec was 160 ± 15°C compared to a Tm, dec of 181 °C for artemisinin It is presumed that thermal decomposition of these tnoxolanes was initiated by formation of a 1,5 diradical produced by homolytic cleavage of the peroxide bond of the tnoxolane nng
Since most of the target tnoxolanes contain the symmetncal spiroadamantane structural framework, their stereochemistry is largely a function of the starting matenal ketone structure or reagents used in post-ozonolysis reactions For OZ27 and other similarly 1,4-substituted tnoxolanes, two achiral diastereomers are possible However, as exemplified by OZ27, the majonty of these tnoxolanes were isolated as single achiral diastereomers rather than as mixtures of two achiral diastereomers For example, in OZ27, no chirahty is present since the tnoxolane nng and phenyl substituent are in a 1,4 relationship in a six membered nng Such compounds possess a plane of symmetry
(Formula Removed)
As determined by X-ray crystallography, the assignment of stereochemistry for OZ78, OZ209 and their denvatives was determined to be cis where the peroxide oxygens are in an axial position
The starting matenal 2-adamantanone may be obtained from Aldnch Chemical Co or from TCI Amencan Organic Chemicals or may also be synthesized Persons skilled in the art can readily ascertain other appropnate means of synthesizing the starting matenals and compounds in accordance with this invention
The spiro and dispiro tnoxolane compositions of the present invention may be generally used for the prophylaxis and treatment of malaria. The tnoxolane compositions of the present invention are administered along with a pharmaceutically acceptable earner Any pharmaceutically acceptable earner may be generally used for this purpose, provided that the earner does not significantly interfere with the stability or bioavailability of the tnoxolane compounds of this invention
The tnoxolanes of this invention can be administered in any effectively pharmaceutically acceptable form to warm blooded animals, including human and other animal subjects, e g in topical, lavage, oral, suppository, parenteral, or infusible dosage forms, as a topical, buccal, sublingual, or nasal spray or in any other manner effective to deliver the agents The route of administration will preferably be designed to optimize delivery and/or localization of the agents to target cells
In addition to the active compounds 1 e the tnoxolanes, the pharmaceutical compositions of this invention may contain suitable excipients and auxihanes which facilitate processing of the active compounds into preparations which can be used pharmaceutically Oral dosage forms encompass tablets, capsules, and granules Preparations which can be administered rectally include suppositones Other dosage forms include suitable solutions for administration parenterally or orally, and compositions which can be administered buccally or subhngually
The pharmaceutical preparations of the present invention are manufactured in a manner which is itself well known m the art For example the pharmaceutical preparations may be made by means of conventional mixing, granulating, dragee-makmg, dissolving, lyophihzing processes. The processes to be used will depend ultimately on the physical properties of the active ingredient used
Suitable excipients are, in particular, fillers such as sugars for example, lactose or sucrose manmtol or sorbitol, cellulose preparations and/or calcium phosphates, for example, tricalcium phosphate or calcium hydrogen phosphate, as well as binders such as starch, paste, using, for example, maize starch, wheat starch, nee starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/or polyvinyl pyrrohdone If desired, disintegrating agents may be added, such as the above-mentioned starches as well as carboxymethyl starch,
cross-linked polyvinyl pyrrohdone, agar, or alginic acid or a salt thereof, such as sodium alginate Auxiliaries are flow-regulating agents and lubricants, for example, such as silica, talc, stearic acid or salts thereof, such as magnesium stearate or calcium stearate and/or polyethylene glycol Oral dosage forms may be provided with suitable coatings which, if desired, may be resistant to gastnc juices
For this purpose concentrated sugar solutions may be used, which may optionally contain gum arable, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures In order to produce coatings resistant to gastnc juices, solutions of suitable cellulose preparations such as acetylcellulose phthalate or hydroxypropylmethylcellulose phthalate, dyestuffs and pigments may be added to the tablet coatings, for example, for identification or in order to charactenze different combination of compound doses.
Other pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol The push-fit capsules can contain the active compounds in the form of granules which may be mixed with fillers such as lactose, binders such as starches, and/or lubneants such as talc or magnesium stearate and, optionally, stabilizers In soft capsules, the active compounds are preferably dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition stabilizers may be added Possible pharmaceutical preparations which can be used rectally include, for example, suppositones, which consist of a combination of the active compounds with the suppository base Suitable suppository bases are, for example, natural or synthetic tnglycendes, paraffin hydrocarbons, polyethylene glycols, or higher alkanols In addition, it is also possible to use gelatin rectal capsules which consist of a combination of the active compounds with a base Possible base matenal include for example liquid tnglycendes, polyethylene glycols, or paraffin hydrocarbons
Suitable formulations for parenteral administration include aqueous solutions of active compounds in water-soluble or water-dispersible form In addition, suspensions of the active compounds as appropnate oily injection suspensions may be administered Suitable lipophilic solvents or vehicles include fatty oils for example, sesame oil, or synthetic fatty acid esters, for example, ethyl oleate or tnglycendes Aqueous injection
suspensions may contain substances which increase the viscosity of the suspension, including for example, sodium carboxymethyl cellulose, sorbitol and/or dextran Such compositions may also comprise adjuvants such as preserving, wetting, emulsifying, and dispensing agents They may also be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating stenlizing agents into the compositions They can also be manufactured in the form of sterile solid compositions which can be dissolved or suspended in sterile water, saline, or other injectable medium poor to administration
In addition to administration with conventional earners, active ingredients may be administered by a variety of specialized delivery drug techniques which are known to those of skill m the art, such as portable infusion pumps
The tnoxolane compositions of the present mvention are administered along with a pharmaceutically acceptable earner in an amount sufficient to prevent malanal infection and/or treat an active infection. The tnoxolane compounds of this invention have extremely low toxicity and a low degree of side effects even at high doses The dosing range of the tnoxolane compositions will vary depending on a number of factors, such as whether it is used for prophylaxis or treatment of an active infection, route of administration, dosing schedule, etc In general, the therapeutic dose of tnoxolane may range between about 0 1-1000 mg/kg/day, with between about 1-100 mg/kg/day being prefened The foregoing doses may be administered as a single dose or may be divided into multiple doses for administration The tnoxolane compositions may be administered once to several times daily For malana prevention, a typical dosing schedule could be, for example, 2 0-1000 mg/kg weekly beginning 1-2 weeks pnor to malana exposure taken up until 1-2 weeks post-exposure
The spiro and dispiro tnoxolanes of this invention may be administered as any pharmaceutically effective salt form Such salts are well known in the art and include, but are not limited to acetate, adipate, alginate, citrate, aspartate, benzoate, benzene sulfonate, bisulfate, butyrate, camphorate, camphor sulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrochlonde, hydrobromide, hydroiodide, 2-hydroxyethansulfonate (isothionate), lactate, maleate, methane sulfonate, nicotmate, 2-naphthalene sulfonate, oxalate, palmitoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, phosphate, glutamate,
bicarbonate, p-toluene sulfonate and undecanoate salts. Preferred salts are those that increase the bioavailability of the tnoxolane compounds. This will depend upon a number of factors, including the chemical structure of the tnoxolane, the carrier to which it is incorporated, the route of administration, etc.
The spiro and dispiro tnoxolanes of this invention have been found to be effective in the treatment of schistosomiasis Schistosomiasis ranks second behind malana m terms of socioeconomic and public health importance m tropical and subtropical areas The disease is endemic m 74 developing countnes, infecting more than 200 million people in rural agricultural and pen-urban areas. An estimated 500-600 million people worldwide are at nsk from the disease
The major forms of human schistosomiasis are caused by five species of water-borne flatworm, or blood flukes, called schistosomes One of these species is Schistosoma mansoni, which has been reported in 53 countnes in Africa, the Eastern Mediterranean, the Canbbean, and South Amenca. The parasites enter the body through contact with infested surface water, pnmanly among people engaged in agnculture and fishing The parasites normally infect the host dunng the cercaria, or larval stage. Once mside the host, the cercana develop into adults or schistosomes
Current treatments for schistosomiasis have focused pnmanly on prophylaxis, 1 e prevention of host infection by cercana Currently, praziquantel is the most widely used drug for treatment of schistosomiasis. While artemether has demonstrated activity m the prophylaxis of schistosomiasis, it has not shown any activity against adult S mansoni
It has now been unexpectedly discovered that the spiro and dispiro tnoxolanes of this invention are active against both cercana and adult S mansoni, S japonwum when administered in the dosages and manner outlined above with respect to treatment of malanal parasites It is also believed the tnoxolanes of this invention will be active against S haematobium Preferred compounds identified for use in the treatment of schistosomiasis include OZ05, OZ11, OZ23, OZ25, OZ28, OZ32, OZ71, OZ78, OZ89, OZ90, OZ119, OZ145, OZ179, OZ205, OZ207, and OZ209 Most preferred compounds are OZ78, OZ207, and OZ209. Preferred dosing levels of the spiro and dispiro tnoxolanes are about 100-200 mg/kg/day orally The prototype tnoxolanes of this invention are OZ03 and OZ05

The spiro and dispiro trioxolanes of this invention may also have effectiveness in treatment of cancer. Compounds having an endoperoxide moiety that is reactive with heme and iron have shown an ability to kill cancer cells. (See e.g. U.S. Pat No. 5,578,637, disclosure of which is hereby incorporated by reference). As noted with respect to artemisinin, trioxolanes' mechanism of action against malarial parasites is based on the ability of trioxolahe compounds to react with the iron in free heme molecules in malaria parasites, with the generation of free radicals leading to cellular destruction. Similarly, trioxolanes are selective against cancer cells due to the higher concentration of transferrin eceptors on cancer cellmembranes that pick up iron at a higher rate than normal cells. In the presence of the trioxolanes of this invention, the cancer cells will accumulate high concentrations of free radicals, leading to cell death. For cancer treatment, the trioxolanes of this invention may be administered in the doses and manner outlined above.
Other drugs besides trioxolanes which are compatible with the carrier ingredients may also be incorporated into the carrier. Such drugs may be readily ascertained by those of ordinary skill in the art and may include, for instance, antibiotics, other antimalarials, antiinflammatory agents, etc.
It is understood that the present invention contemplates the use of not only the above-stated trioxolane compounds themselves, but their prodrugs which metabolize to the compound and the analogues and biologically active salt forms thereof, as well as optical isomers which provide the same pharmaceutical results.
The following examples are offered to illustrate but not limit the invention. Thus, they are presented with the understanding that various formulation modifications as well as method of delivery modifications may be made and still be within the spirit of the invention.
The present invention relates to a spiro or dispiro 1,2,4-trioxoiane substantially as hereinbefore described with reference to the accompanying drawings.
The present invention further relates to a pharmaceutical composition for prophylaxis and treatment of malaria substantially as hereinbefore described with reference to the accompanying drawings.
EXAMPLE 1 Antimalarial Activity of OZ271-OZ369
Antimalarial Assays
Various OZ compounds were tested by the semiautomated microdilution assay against intraerythrocytic forms of Plasmodium falciparum derived from asynchronous stock cultures. The culture medium used was RPMI1640 supplemented with 10% human

type A+ serum, 25 mM HEPES, 25 mM NaHC03 (pH 7.3). Human type A+ erythrocytes served as host cells The culture was kept at 37°C in an atmosphere of 3% 02,4% C02, and 93% N2 in humidified modular chambers.
Compounds were dissolved in DMSO (10 mg/ml), pre-diluted in complete culture medium, and titrated in duplicate in senal twofold dilutions over a 64-fold range in 96-well microtiter plates. After addition of the parasite cultures with an initial parasitemia (expressed as the percentage of erythrocytes infected) of 0.75% in a 2 5% erythrocytes suspension, the test plates were incubated under the conditions described above for 72 h Growth of the parasites cultures was measured by the incorporation of radiolabeled [3H]-hypoxanthme added 16 h prior to termination of the test. Fifty percent inhibitory concentration (IC50) were estimated by Logit regression analysis Compounds were tested against reference P falciparum strains, Kl strain (Thailand resistant to chloroquine) and NF54 strain (an airport strain of unknown origin that is sensitive to standard antimalarials)
In the single dose STI in vivo screen, Moro SPF or NMRI mice infected with the ANKA strain of P berghei (groups of 3-5 mice) were used Tnoxolanes were administered as single oral (po) 3 mg/kg and 10 mg/kg doses in standard suspending vehicle (SSV) SSV consists of 0.5% w/v CMC, 0.5% v/v benzyl alcohol, 0 4% v/v Tween 80, and 0.9% w/v sodium chlonde in water Antimalanal activity was measured by percent reduction in parasitemia on day three post-infection and survival times compared to an untreated control group. Survival to day 30 post-infection is considered to be a cure. In PCT/02/19767, Table 1 presented data for tnoxolanes OZ01-OZ270 along with the controls, fenozan, artemisinin, arteether, artemether, and artesunate The data showed that antimalanal activity falls off both when the tnoxolane peroxide bond is too exposed or is stencally inaccessible to iron(II) species Other factors influencing antimalanal activity include the stability of carbon radicals formed by J3-scission subsequent to the initial electron transfer to the peroxide bond and the influence of stenc effects remote from the peroxide bond on the interactions between carbon radicals and potential drug targets The data also demonstrated that tnoxolane carboxyhc acids are usually less active than their hydrocarbon, ester, and hydroxamic acid counterparts
Below is the activity data for OZ271-OZ369.
(Table Removed) …………..1
For comparative analysis, data is also presented for the control antimalarial drugs artemether (AM), artesunate (AS), chloroqume (CQ), and mefloquine (MFQ)
The new activity data demonstrates that tnoxolane carboxyhc acids are usually less active than their hydrocarbon, ester, amide, and hydroxamic acid counterparts The position of lonizable functional groups such as carboxyhc acids and amines is also critical to activity The best combination of high intrinsic potency and good oral activity is found when a weak base functional group is present
Onset of Action and Recrudescence of OZ11, OZ27, OZ78, OZ156, OZ175, OZ177, OZ207, OZ209, OZ277, and OZ279
Onset of Action and Recrudescence Expenments
The onset of drug action was determined after a single fixed dose of 100 mg/kg (SSV vehicle) po to groups of five animals on day +3 post- infection (day 0) Parasitemias
at this point are usually between 25-40% The infected controls do not survive beyond day +6 post-infection The reduction of parasitemia is monitored 12,24, and 48 h after treatment, and the time of recrudescence (> 5% parasitemia) is assessed by daily blood smears for 14 days, followed by intermittent assessment for up to 60 days
The onset part of this experiment reveals how rapidly a compound reduces parasite load; the recrudescence part of the experiment provides information about the efficacy of the compound against the parasite A long delay in recrudescence can be due to a very good antiparasitic effect of the compound or to a compound with a long half-life
Both the tnoxolanes and the artemismins produced a rapid decline in parasitemia, confirming that they are rapidly acting antimalanal agents. In contrast to both chloroquine and these peroxidic antimalarials, mefloquine has a slow onset of action Recrudescence (> 5% parasitemia) occurs quite rapidly for artemisinin and artesunate The time of recrudescence increased for the more lipophilic artemisinin derivatives artemether and arteether
hi contrast to artemether, recrudescence occured much more slowly for the lipophilic tnoxolanes OZ11 and OZ27, the recrudescence time for OZ27 was especially marked, supenor to that of mefloquine However, recrudescence times for the relatively polar tnoxolanes OZ78, OZ175, and OZ277 were very similar to that of artemether The more lipophilic tnoxolane (OZ156) of the OZ156/OZ177 pair produced the longest delay in recrudescence, longer than chloroquine, but less than mefloquine The recrudescence times for OZ177 and OZ279 were roughly equivalent to that of chloroquine
Stnkmgly, there was no recrudescence observed for OZ207 and OZ209, two different salt forms (OZ207 - tosylate, OZ209 - mesylate) of ammomethyl tnoxolane OZ163 (hydrochlonde) The recrudescence data for these two tnoxolanes suggests that they are either more powerful antimalanal agents or have longer half-lives than any of the semisynthetic artemismins.
Table 2
(Table Removed)
EXAMPLE 3 Effect of Trioxolanes on Schistosoma Species
Effect of Tnoxolane OZ207 on Schistosoma laponicum
Table 3 Comparative effect of OZ207 and artemether in mice infected with Schistosoma laponicum
(Table Removed)
MTWB, mean total worm burden; WRR, worm reduction rate MFWB, mean female worm burden, FWRR, female worm reduction rate
Table 3 illustrates that the mean total worm burden and mean female worm burden in OZ207 400 mg/kg group was significantly lower than those in artemether 400 mg/kg group (P Mice were infected with 100 Schistosoma mansoni cercanae on day 21 post-treatment Each group was treated per os with tnoxolanes at a single dose of 200 mg/kg Untreated mice served as the control All groups were killed 4 weeks after treatment and the liver and intestine were removed and separated. The liver and intestine were compressed and alive male and female worms could be seen and counted The effect of the compounds was evaluated by mean total and female worm burden The results are shown m Table 4 Effect of Tnoxolanes on adult schistosomes (49-dav-old)
Mice were infected with 100 Schistosoma mansoni cercanae on day 49 post-treatment Each group was treated per os with OZ compounds at single doses of 200,400, and 600 mg/kg Untreated mice served as the control All groups were killed 4 weeks after treatment and the liver and intestine were removed and separated The liver and intestine were compressed and alive male and female worms could be seen and counted The effect of the compounds was evaluated by mean total and female worm burden, and the results are set forth in Table 4
Table 4
(Table Removed)
EXAMPLE 4 Activity of Trioxolanes Against P. berghei
In the single dose ED50/ED90/ED99 determinations, Moro SPF or NMRI mice (group of three) infected with the ANKA strain of Plasmodium berghei were treated on day one post-infection Tnoxolanes were dissolved or suspended in the standard suspending vehicle (SSV) and administered as single 10, 6, 3,1, 0 3, and 0 1 mg/kg doses po and sc The SSV consists of 0 5% w/v CMC, 0 5% v/v benzyl alcohol, 0 4% v/v Tween 80, and 0 9% w/v sodium chloride in water. Antimalarial activity was measured by percent reduction in parasitemia on day three post-infection. The ED50/ED90 values were calculated by nonlinear fitting
Table 5
(Table Removed)
Table 5 shows ED50/ED90/ED99 data obtained by po administration of tnoxolanes in the SSV formulation The relatively lipophilic artemether is substantially more active than the more polar artesunate and artelmate In contrast, the most active tnoxolanes (OZ181, OZ207, OZ209) - different salt forms of the same ammo tnoxolane, and ammo and amide tnoxolanes OZ277 and OZ279, are relatively polar compounds
EXAMPLE 5 Dosing of OZ279, OZ277, OZ256, and OZ209
Based on results of dosing OZ279, OZ277, OZ256, and OZ209 in rats and dogs, the inventors determined projected optimal dosing of the same compounds in humans Artesunate is listed as a reference compound
Table 6
(Table Removed)
Effectiveness of Selected OZ Compounds in the Treatment and Prophylaxis of Malarial Infections
Moro NMRI male mice (Fu Albino specific pathogen free) weighing 18 ± 2 g were infected intravenously (l v.) with 2 x 107 P berghei ANKA strain-mfected erythrocytes from donor mice on day 0 of the experiment From donor mice with circa 30% parasitemia, hepanmzed blood was taken and diluted in physiological saline to 10 parasitized erythrocytes per ml An aliquot (0 2 ml) of this suspension was injected l v into experimental and control groups of mice. In untreated control mice, parasitemia rose regularly to 40 to 50% by day 3 post-infection and 70 to 80% by day 4 post-infection The mice died between days 5 and 7 post-infection Throughout the experiments, mice were kept m groups of three or five animals in Makrolon type II cages in an air-conditioned animal room at 22 to 23°C A diet withp-aminobenzoic acid (PABA) of 45 mg (NAFAG FUTTER® food N° 9009 PAB-45) per kg of body weight, and tap water is available ad libitum
OZ compounds were prepared at an appropriate concentration, either as a solution or a suspension containing SSV (0 5% w/v CMC, 0 5% v/v benzyl alcohol, 0 4% v/v Tween 80, and 0 9% w/v sodium chloride in water) They were administered per os (p o ) in a total volume of 0 01 ml per gram of mouse. The activity of the compound was
determined by a vanety of methods outlined in subsequent sections Survival time was also recorded, and survival to day 30 post-infection was considered to be a cure
The first experiment conducted consisted of administration of a divided 3x10 mg/kg p o dose administered on days 1, 2, and 3 post-infection vs a single 1 x 30 mg/kg po dose administered on day 1 post-infection On day 4 post-infection, blood smears of all animals were prepared and stained with Giemsa Parasitemia was determined microscopically, and the difference between the mean value of the control group (taken as 100%) and those of the experimental groups was calculated and expressed as percent reduction. Compounds were administered orally in the SSV vehicle The results are shown in Table 7 below:
Table 7

(Table Removed)
As shown by Table 7, a 3 x 10 mg/kg dose of these tnoxolanes cured between 3/5 and 5/5 of the infected mice At this same dose, none of the standard antimalarial drugs cured any of the infected mice At the 1 x 30 mg/kg dose, all tested tnoxolanes showed activities > 99 9% on day 3 post-treatment
The second expenment consists of administration of divided 3x3 mg/kg and 3 x 1
mg/kg po doses administered on days 1, 2, and 3 post-infection On day 4 post-infection,
blood smears of all animals were prepared and stained with Giemsa Parasitemia was
determined microscopically, and the difference between the mean value of the control
group (taken as 100%) and those of the expenmental groups was calculated and expressed as percent reduction Compounds were administered orally in the SSV vehicle The results are shown in Table 8.
(Table Removed)
As shown by Table 8, at the 3 x 3 mg/kg dose, fourteen tnoxolanes had activities of 100% and produced high survival numbers Of these, OZ301, OZ329, OZ339, OZ349, and OZ357 cured 1/5, 2/5, 3/5, 2/5, and 1/5 of the infected mice, respectively At the 3 x 1 mg/kg dose, most of the tnoxolanes were more potent than the reference antimalanal drugs; sixteen of these had activities > 90% OZ209, OZ329, and OZ336 were the only tnoxolanes with activities greater than 99% at the 3 x 1 mg/kg dose All of OZ343-OZ368 that were tested were more active than the reference antimalanal drugs
Prophylactic activities of the compounds were compared after admmistenng po single dose of 100 mg/kg to different groups of five animals at vanous times before infection All groups including an untreated control group, were then infected at the same time Parasitemia was determined for each animal on day 3 post-infection, and percent of reduction of the level of parasitemia compared to levels for animals given no drug is determined The results are shown in Table 9
(Table Removed)
The unique prophylactic property of OZ209 (3-day protection, same as MFQ) was found also for OZ271
EXAMPLE 7 Chemical Properties of OZ277 Salts
The following tosylate, maleate, hydrochloride, tartarate, and succinate salts of the base form of OZ277 were synthesized
cis-Adamantane-2-spiro-3,-8,-[[[(2'-amino-2,-methylpropyl)ammo]carbonyl]methyl]-1,2',4'-tnoxaspiro[4 5]decane/?-tosylate
cis-Adamantane-2-spiro-3'-8'-[[[(2,-amino-2'-methylpropyl)ammo]carbonyl]methyl]-1,2',4'-tnoxaspiro[4 5]decane hydrogen maleate
cis-Adamantane-2-spiro-3'-8'-[[[(2'-amino-2'-methylpropyl)amino]carbonyl]methyl]-1,2',4'-tnoxaspiro[4 5]decane hydrochloride
cis-Adamantane-2-spiro-3'-8'-[[[(2'-amino-2'-methylpropyl)ammo]carbonyl]methyl]-1,2',4'-tnoxaspiro[4 5]decane hydrogen tartarate
cis-Adamantane-2-spiro-3'-8'-[[[(2'-amino-2'-methylpropyl)amino]carbonyl]methyl]-1,2',4'-tnoxaspiro[4 5]decane hydrogen succinate
Their properties are shown in Table 10:
(Table Removed)
EXAMPLE 8 Preparation of OZ277 Maleate Salt
Raw Materials
(Table Removed)
-Charge ethanol (150 ml) at 20-25°C,
-Charge OZ277 (60 gm) at 20-25 °C under stirring;
-Stir at 20-25°C for 10 mm to get clear solution,
-Add the solution of maleic acid (17 4 g in 90 ml ethanol) drop wise at 20-25°C under stirring in 1 hr ,
-Charge Heptane (720 ml) at 20-25°C under stirring,
-Stir for 4 hr at 20-25°C,
-Filter the solid and wash with heptane ( 60 ml),
-Suck dry the product,
-Dry the product under vacuum at 20-25 °C
The resulting product has a weight of 65-70 grams.
It should be appreciated that the spiro and dispiro 1,2,4-tnoxolane compositions of this invention may contain tnoxolanes within the scope of the formulas descnbed above, or prodrugs or analogues of these compounds or a racemic mixture of either the D or the L form The invention is also intended to include all biologically active salt forms of the compounds Also, minor dosage and formulation modifications of the composition and the ranges expressed herein may be made and still come within the scope and spint of the present invention
Having descnbed the invention with reference to particular compositions, theones of effectiveness, and the like, it will be apparent to those of skill in the art that it is not intended that the invention be limited by such illustrative embodiments or mechanisms, and that modifications can be made without departing from the scope or spint of the invention, as defined by the appended claims It is intended that all such obvious modifications and vanations be included within the scope of the present invention as defined in the appended claims The claims are meant to cover the claimed components and steps in any sequence which is effective to meet the objectives there intended, unless the context specifically indicates to the contrary.

We claim :
1. A spiro or dispiro 1,2,4-trioxolane having the following structural formula:
(Formula Removed)
whereby R3 is (CH2)n-Y and Y represents a weak base, wherein the spiro or dispiro 1,2,4-trioxolane is selected from the group consisting of:
(Formula Removed)
3is-Adamantane-2-spiro-3'-8'-[[(aminoacetyl)amino]methyl]-1',2',4'-trioxaspiro [4.5]decane mesylate;
(Formula Removed)
(Formula Removed)
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(2,-amino-2'-methylpropyl)amino]carbonyl] methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[(1'-piperazinylcarbonyl)methyl]-1',2',4'-trioxaspiro [4.5]decane p-tosylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[(2'-hydroxybenzoyl)amino]methyl]-1',2',4'-trioxaspiro [4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8,-[[(2,-amino-2'-methylpropionyl)amino] methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(4,-hydroxy-3'-pyridinyl)carbonyl]amino] methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(3'-amino-1'H-triazol-5'-yl)carbonyl] amino]methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(2'-amino-3'-pyridinyl)carbonyl] amino]methyl]-1',2',4'-trioxaspiro[4.5]decane mesylate;
(Formula Removed)
(Formula Removed)
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(2'-pyridinylmethyl)amino]carbonyl]methyl]-1',2',4'-trioxaspiro[4.5]decane mesylate;
(Formula Removed)
(Formula Removed)
(Formula Removed)
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'[[(2'-pyridinylcarbonyl)amino]methyl]-1',2',4'-trioxaspiro [4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8,-[[[(2'-aminoethyl)amino]carbonyl]methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8' -[[[(2'-amino-2'-oxoethyl)amino]carbonyl] methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[4'-(aminocarbonyl)-1 '-piperidinyl] carbonyl]methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[(4'-carboxy-1'-piperidinyl)carbonyl] methyl] 1',2',4'-trioxaspiro[4.5]decane;-
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[[2'-(4,-morpholinyl)ethyl]amino] carbonyl] methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[4'-(2'-pyrimidinyl)-1'-piperazinyl]carbonyl] methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'[[[trans-4'-aminocyclohexyl) amino]carbonyl]methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[2'-[(3'-pyridinylcarbonyl)amino] acetyl]amino] methyl]-1',2',4'-trioxaspiro[4.5]decane mesylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(1 '-aminocyclopentyl)carbonyl] amino]methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[(2'S)-2'-aminocarbonyl-1'-pyrrolidinyl] methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[(1 '-piperazinylcarbonyl)amino]methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(3'-amino-2'-pyrazinyl)carbonyl]amino] methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
(Formula Removed)
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[(3'-aminopropionyl)amino]methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(2'S)-2'-pyrrolidinylcarbonyl]amino] methyl]-1',2',4'-trioxaspiro[4.5]decane;

(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(3'S)-3'-amino-1'-pyrrolidinyl]carbonyl] methyl]1',2',4'-trioxaspiro[4.5]decane mesylate;
(Formula Removed)
piperidinyl)carbonyl] methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-(2,-oxo-2'-guanidinoethyl)-1',2',4'-trioxaspiro[4.5] decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[(1', 1 '-dioxido-4'-thiomorpholinyl)carbonyl] methyl]-1',2',4'-trioxaspiro [4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(3'-amino-2',2'-dimethylpropyl)amino]carbonyl]methyl]-1',2',4'-trioxaspiro[4.5]decane mesylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[4'-(methylsulfonyl)-1'-piperazinyl] carbonyl]methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[4,-(methylsulfonyl)-1 -piperidinyl] carbonyl]methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8,-[[[(2'S)-2'-aminocarbonyl-1'-pyrrolidinyl] carbonyl]methyl]-1',2',4'-trioxaspiro[4.5]decane;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[[(trans-4'-aminocyclohexyl) amino]carbonyl]amino]methyl]-1',2',4'-trioxaspiro[4.5]decane
(Formula Removed)
cis-Adamantane-2-spiro-3,-8'-[[[[(2,-amino-2'-methylpropyl)amino] carbonyl]amino]methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
N-(cis-Adamantane-2-spirc~3'-1',2',4'-trioxaspiro[4.5]decane-8'-methyl)-N'-(trans-4-aminocyclohexyl)oxamide p-tosylate;
(Formula Removed)
(Formula Removed)
N-(cis-Adamantane-2-spirc-3'-1',2',4'-trioxaspiro[4.5]decane-8'-methyl)-N'-(2-amino-2-methylpropyl)oxamide mesylate;
(Formula Removed)
(Formula Removed)
N-(cis-Adamantane-2-spiro-3'-1',2',4'-trioxaspiro[4.5]decane-8'-methyl)-N'-(4-piperidinyl)oxamide mesylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[[1'-aminocyclopropyl)methyl]amino] carbonyl] methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
N-[(cis-Adamantane-2-spiro-3'-1',2,,4,-trioxaspiro[4.5]decan-8'-yl)methyl] oxamic acid;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(3'-aminopropyl)amino]carbonyl]methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(2,-amino-2'-methylpropyl)amino] carbonyl] amino]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
trans-Adamantane-2-spiro-3'-8'-(1 '-piperazinylcarbonyl)-
1',2',4'-trioxaspiro[4.5] decane p-tosylate;
(Formula Removed)
trans-Adamantane-2-spiro-3'-8'-[[(2'amino-2'-methylpropyl)amino] carbonyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8,-[[(2'-amino-2'-methylpropyl)amino] carbonyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-(1 '-piperazinylcarbonyl)-1',2',4'-trioxaspiro [4.5] decane p-tosylate; and
(Formula Removed)
N-(cis-Adamantane-2-spiro-3'-1',2',4'-trioxaspiro[4.5]decane-8'-methyl)-a-oxo-1-piperazineacetamide mesylate; cis-adamantane-2-spiro-3'-8'-[[[(2'-amino-2'-methylpropyl)amino]carbonyl]methyl]-1',2',4'-trioxaspiro [4.5]decane; cis-adamantane-2-spiro-3'-8'-[[[(2'-amino-2'-
methylpropyl)amino]carbonyl]methyl]-1',2',4'-trioxaspiro [4.5]decane hydrogen maleate;
cis-adamantane-2-spiro-3'-8'[[[(2'-amino-2'-methylpropyl)amino]carbonyl]methyl]-1',2',4'-trioxaspiro [4.5]decane hydrochloride;
methylpropyl)amino]carbonyl]methyl]-1',2',4'-trioxaspiro [4.5] decane hydrogen tartrate, and cis-adamantane-2-spiro-3'-8'-[([(2'-amino-2,-methylpropyl)amino] carbonyl]methyl]-1',2',4'-trioxaspiro[4.5]decane hydrogen succinate.
2. The spiro or dispiro 1,2, 4-trioxolane as claimed in claim 1 that is selected from the
consisting of
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[(2'-amino-2'-methylpropyl)amino]carbonyl] methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate;
(Formula Removed)
N-(cis-Adamantane-2-spiro-3'-1',2',4'-trioxaspiro[4.5]decane-8'-methyl)-NXtrans-4-aminocyclohexyl)oxamide p-tosylate;
(Formula Removed)
N-(cis-Adamantane-2-spiro-3'-1',2',4'-trioxaspiro[4.5]decane-8'-methyl)-N'-(2-amino-2-methylpropyl)oxamide mesylate;-
(Formula Removed)
N-(cis-Adamantane-2-spiro-3'-1',2,,4'-trioxaspiro[4.5]decane-8'-methyl)-N'-(4-piperidinyl)oxamide mesylate;
(Formula Removed)
cis-Adamantane-2-spiro-3'-8'-[[[[(1 '-aminocyclopropyl)methyl]amino] carbonyl] methyl]-1',2',4'-trioxaspiro[4.5]decane p-tosylate; and

(Formula Removed)
cis-Adamantane-2-spiro-3'-8,-(1'-piperazinylcarbonyl)-1',2',4'-trioxaspiro[4.5] decane p-tosylate.
3. A compound as claimed in claim 1 wherein the spiro or dispiro 1,2,4-trioxolane is
1',2',4'-trioxaspiro [4.5]decane hydrogen maleate.
4. A spiro or dispiro 1,2,4-trioxolane substantially as hereinbefore described with reference to the accompanying drawings.
















1514-del-2004-Description (Complete)-(26-02-2010).pdf

1514-del-2004-description (complete).pdf











1514-del-2004-Petition 137-(26-02-2010).pdf

Patent Number 245779
Indian Patent Application Number 1514/DEL/2004
PG Journal Number 05/2011
Publication Date 04-Feb-2011
Grant Date 01-Feb-2011
Date of Filing 16-Aug-2004
# Inventor's Name Inventor's Address
PCT International Classification Number A61K 31/4747
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 10/642,721 2003-08-18 U.S.A.