RESUMEN
The tumor microenvironment in chondrosarcoma (CHS), a chemo- and radio-resistant cancer provides unique hallmarks for developing a chondrosarcoma targeted drug-delivery system. Tumor targeting could be achieved using a quaternary ammonium function (QA) as a ligand for aggrecan, the main high negative charged proteoglycan of the extracellular matrix of CHS, and a 2-nitroimidazole as trigger that enables hypoxia-responsive drug release. In a previous work, ICF05016 was identified as efficient proteoglycan-targeting hypoxia-activated prodrug in a human extraskeletal myxoid chondrosarcoma model in mice and a first study of the structure-activity relationship of the QA function and the alkyl linker length was conducted. Here, we report the second part of the study, namely the modification of the nitro-aromatic trigger and the position of the proteoglycan-targeting ligand at the aromatic ring as well as the nature of the alkylating mustard. Synthetic approaches have been established to functionalize the 2-nitroimidazole ring at the N-1 and C-4 positions with a terminal tertiary alkyl amine, and to perform the phosphorylation step namely through the use of an amine borane complex, leading to phosphoramide and isophosphoramide mustards and also to a phosphoramide mustard bearing four 2-chloroethyl chains. In a preliminary study using a reductive chemical activation, QA-conjugates, except the 4-nitrobenzyl one, were showed to undergo efficient cleavage with release of the corresponding mustard. However N,N,N-trimethylpropylaminium tethered to the N-1 or C-4 positions of the imidazole seemed to hamper the enzymatic reduction of the prodrugs and all tested compounds featured moderate selectivity toward hypoxic cells, likely not sufficient for application as hypoxia-activated prodrugs.
Asunto(s)
Antineoplásicos/farmacología , Neoplasias Óseas/tratamiento farmacológico , Condrosarcoma/tratamiento farmacológico , Diseño de Fármacos , Neoplasias de los Tejidos Conjuntivo y Blando/tratamiento farmacológico , Mostazas de Fosforamida/farmacología , Profármacos/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/química , Neoplasias Óseas/patología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Condrosarcoma/patología , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Estructura Molecular , Neoplasias de los Tejidos Conjuntivo y Blando/patología , Mostazas de Fosforamida/síntesis química , Mostazas de Fosforamida/química , Profármacos/síntesis química , Profármacos/química , Relación Estructura-ActividadRESUMEN
A series of Glutaryl-Hyp-Ala-Ser-Chg-Gln-4-aminobenzyl phosphoramide mustard conjugates (1a-e) was designed and synthesized as potential prodrugs for site-specific activation by PSA in prostate cancer cells. All conjugates were found to be substrates of PSA with cleavage occurring between Gln and the para-aminobenzyl (PAB) linker. Structure-activity relationship studies on these conjugates indicated that introduction of electron-withdrawing fluorine(s) on the phenyl ring in the PAB linker uniformly improved the chemical stability of the conjugates while the position of substitution affected differently the self-immolative process of conjugates upon proteolysis. Introduction of a fluorine at ortho position to benzylic phosphoramide as in 1b results in better stability of the conjugate prior to activation while maintaining its antiproliferative activity upon activation by PSA. The conjugate 1b with 2-fluoro substitution was identified as a promising lead for further evaluation and optimization in the development of prostate cancer-targeted prodrugs.
Asunto(s)
Antineoplásicos/química , Diseño de Fármacos , Péptidos/química , Mostazas de Fosforamida/química , Profármacos/química , Antígeno Prostático Específico/metabolismo , Neoplasias de la Próstata/tratamiento farmacológico , Antineoplásicos/síntesis química , Antineoplásicos/metabolismo , Antineoplásicos/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Humanos , Masculino , Péptidos/síntesis química , Péptidos/metabolismo , Péptidos/farmacología , Mostazas de Fosforamida/síntesis química , Mostazas de Fosforamida/metabolismo , Mostazas de Fosforamida/farmacología , Profármacos/síntesis química , Profármacos/metabolismo , Profármacos/farmacología , Neoplasias de la Próstata/metabolismo , Relación Estructura-ActividadRESUMEN
A series of N-mustards, which was conjugated to mono- or bis-naphthalimides with a flexible amine link, were synthesized and evaluated for cytotoxicity against five cancer cell lines (HCT-116, PC-3, U87 MG, Hep G2 and SK-OV-3). Several compounds displayed better activities than the control compound amonafide. Further evaluations by fluorescence spectroscopy studies and DNA-interstrand cross-linking assays revealed that the derivatives showed both alkylating and intercalating properties. Among the derivatives, the bis-naphthalimide N-mustard derivative 11b was found to exhibit the highest cytotoxic activity and DNA cross-linking ability. Both 11b and 7b induce HCT-116 cell apoptosis by S phase arrest.
Asunto(s)
Antineoplásicos Alquilantes/síntesis química , Naftalimidas/síntesis química , Mostazas de Fosforamida/síntesis química , Antineoplásicos Alquilantes/farmacología , Apoptosis/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Células HCT116 , Células Hep G2 , Humanos , Concentración 50 Inhibidora , Naftalimidas/farmacología , Mostazas de Fosforamida/farmacología , Poli(ADP-Ribosa) Polimerasa-1 , Poli(ADP-Ribosa) Polimerasas/metabolismoRESUMEN
A series of new prodrugs: [bis(2-chloroethylamino)phosphoryloxy]methyl acetate, [bis(2-chloroethylamino)phosphoryloxy]methyl pivalate and [bis(2-chloroethylamino)phosphoryloxy]methyl benzoate, was obtained in the reaction of isophosphoramide mustard (iPAM) with the corresponding acyloxymethyl halides. The cytotoxic activity of these new compounds is also shown. All compounds were highly active in the inhibition of cancer cell proliferation against the human lung (A594), prostate (PC-3) and breast (MCF-7) cancer cell lines.
Asunto(s)
Antineoplásicos/síntesis química , Mostazas de Fosforamida/síntesis química , Profármacos/síntesis química , Antineoplásicos/farmacología , Biotransformación , Línea Celular Tumoral , Estabilidad de Medicamentos , Humanos , Mostazas de Fosforamida/farmacología , Profármacos/farmacologíaRESUMEN
Cyclophosphamide, an alkylating agent widely used as anticancer agent, biotransformed in vivo to unstable phosphoramidic mustard and acrolein, where the latter metabolite has been found responsible for hemorrhagic cystitis and renal toxicity. Being one of the most popular strategies to avoid these deleterious effects, prodrug design has been attempted, which can, in addition, enable selective drug targeting. Our efforts to design, synthesize and evaluate the enzymatically activated prodrug phosphorodiamidic mustard as potential candidate for selective chemotherapy in antibody-directed enzyme prodrug therapy or prodrug monotherapy strategies are described. We propose an improved synthesis of prodrug 14, consisting of a galactose moiety, a spacer and a cytotoxic drug and its cytotoxicity has been investigated. The prodrug 14 has been found to be nontoxic (in vitro) which could be a valuable candidate for further development.
Asunto(s)
Antineoplásicos Alquilantes/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Sistemas de Liberación de Medicamentos , Diseño de Fármacos , Galactósidos/farmacología , Lisosomas/metabolismo , Mostazas de Fosforamida/farmacología , Profármacos/farmacología , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/metabolismo , Antineoplásicos Alquilantes/efectos adversos , Antineoplásicos Alquilantes/síntesis química , Antineoplásicos Alquilantes/metabolismo , Supervivencia Celular/efectos de los fármacos , Ciclofosfamida/efectos adversos , Ciclofosfamida/análogos & derivados , Ciclofosfamida/metabolismo , Ciclofosfamida/farmacología , Sistemas de Liberación de Medicamentos/efectos adversos , Estabilidad de Medicamentos , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/metabolismo , Femenino , Galactósidos/efectos adversos , Galactósidos/síntesis química , Galactósidos/metabolismo , Células HeLa , Humanos , Hidrólisis , Concentración 50 Inhibidora , Cinética , Células MCF-7 , Mostazas de Fosforamida/efectos adversos , Mostazas de Fosforamida/síntesis química , Mostazas de Fosforamida/metabolismo , Profármacos/efectos adversos , Profármacos/síntesis química , Profármacos/metabolismo , beta-Galactosidasa/química , beta-Galactosidasa/metabolismoRESUMEN
A series of novel compounds with phosphoramide mustard functionality incorporated into the quinazoline scaffold of EGFR/HER2 inhibitors were designed and synthesized as multi-target-directed ligands against tumor cells. In vitro assays showed that tumor cell lines with high HER2 level were more sensitive to the compounds than tumor cells with low HER2 level. Compound 10d (EMB-3) was one of the most potent inhibitors with IC50 of 7.4 nM and 82 nM against EGFR and HER2, respectively. The mechanism studies were also supported by the effect of 10d-induced DNA damage in MDA-MB-468 cells. In vivo efficacy study showed that 10d could significantly inhibit H522 tumor xenograft model with a TGI of 68% at dose of 100 mg/kg (QDx28, p.o.) and no significant body weight loss was observed. MTD study indicated that compound 10d had no acute toxicity to mice at doses up to 900 mg/kg (single dose).
Asunto(s)
Antineoplásicos/síntesis química , Antineoplásicos/farmacología , Diseño de Fármacos , Mostazas de Fosforamida/síntesis química , Mostazas de Fosforamida/farmacología , Quinazolinas/química , Animales , Antineoplásicos/química , Antineoplásicos/farmacocinética , Línea Celular Tumoral , Técnicas de Química Sintética , Relación Dosis-Respuesta a Droga , Humanos , Masculino , Ratones , Mostazas de Fosforamida/química , Mostazas de Fosforamida/farmacocinética , Ratas , Relación Estructura-Actividad , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
In efforts to obtain anticancer prodrugs for antibody-directed or gene-directed enzyme prodrug therapy using E. coli nitroreductase, a series of nitrobenzylphosphoramide mustards were designed and synthesized incorporating a strategically placed nitro group in a position para to the benzylic carbon for reductive activation. All analogues were good substrates of E. coli nitroreductase with half-lives between 2.9 and 11.9 min at pH 7.0 and 37 degrees C. Isomers of the 4-nitrophenylcyclophosphamide analogues 3 and 5 with a benzylic oxygen para to the nitro group showed potent selective cytotoxicity in nitroreductase (NTR) expressing cells, while analogues 4 and 6 with a benzylic nitrogen para to the nitro group showed little selective cytotoxicity despite their good substrate activity. These results suggest that good substrate activity and the benzylic oxygen are both required for reductive activation of 4-nitrophenylcyclophosphamide analogues by E. coli nitroreductase. Isomers of analogue 3 showed 23,000-29,000x selective cytotoxicity toward NTR-expressing V79 cells with an IC(50) as low as 27 nM. They are about as active as and 3-4x more selective than 5-aziridinyl-2,4-dinitrobenzamide (CB1954). The acyclic 4-nitrobenzylphosphoramide mustard ((+/-)-7) was found to be the most active and most selective compound for activation by NTR with 170,000x selective cytotoxicity toward NTR-expressing V79 cells and an IC(50) of 0.4 nM. Compound (+/-)-7also exhibited good bystander effect compared to 5-aziridinyl-2,4-dinitrobenzamide. The low IC(50), high selectivity, and good bystander effects of nitrobenzylphosphoramide mustards in NTR-expressing cells suggest that they could be used in combination with E. coli nitroreductase in enzyme prodrug therapy.
Asunto(s)
Antineoplásicos/síntesis química , Proteínas de Escherichia coli/metabolismo , Nitrorreductasas/metabolismo , Mostazas de Fosforamida/síntesis química , Profármacos/síntesis química , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Línea Celular , Línea Celular Tumoral , Cricetinae , Cricetulus , Diseño de Fármacos , Activación Enzimática , Proteínas de Escherichia coli/genética , Humanos , Nitrorreductasas/genética , Mostazas de Fosforamida/química , Mostazas de Fosforamida/farmacología , Profármacos/química , Profármacos/farmacología , Estereoisomerismo , Relación Estructura-Actividad , TransfecciónRESUMEN
Aldophosphamide, the penultimate cytotoxic metabolite of cyclophosphamide, can be detoxified by an oxidation reaction catalyzed by certain aldehyde dehydrogenases. The selective toxicity of cyclophosphamide is due, at least in part, to a greater expression of the relevant aldehyde dehydrogenase activity in normal cells relative to that expressed in certain tumor cells. Not known at the onset of this investigation was which of the several known mouse aldehyde dehydrogenases catalyze this reaction. Twelve enzymes that catalyze the NAD(P)-linked oxidation of aldophosphamide, acetaldehyde, benzaldehyde, and/or octanal were chromatographically resolved from mouse liver. Four of these appear to be novel; four others were determined to be betaine aldehyde dehydrogenase, succinic semialdehyde dehydrogenase, glutamic gamma-semialdehyde dehydrogenase, and xanthine oxidase (dehydrogenase). An additional aldehyde dehydrogenase, namely AHD-4, was semipurified from stomach. The stomach enzyme and nine of the hepatic enzymes catalyze the oxidation of aldophosphamide. Km values for these reactions range from 16 microM to 2.5 mM. The relevant aldehyde dehydrogenase of major importance varies with the tissue. In the liver, the major cytosolic aldehyde dehydrogenase, namely AHD-2, accounts for greater than 60% of total hepatic aldehyde dehydrogenase-catalyzed aldophosphamide (160 microM) detoxification. Succinic semialdehyde dehydrogenase (AHD-12) and three of the novel hepatic aldehyde dehydrogenases, namely AHD-8, AHD-10, and AHD-13, also contribute significantly to total hepatic aldehyde dehydrogenase-catalyzed aldophosphamide detoxification. In the stomach, AHD-4 and AHD-8 account for approximately 86% of total aldehyde dehydrogenase-catalyzed aldophosphamide (160 microM) detoxification. AHD-2 was not found in this tissue. Of all the aldehyde dehydrogenases examined, AHD-2 and AHD-8 were estimated to be the most efficient catalysts of aldophosphamide oxidation. Thus, these enzymes would seem most likely to be operative when tumor cells acquire aldehyde dehydrogenase-mediated cyclophosphamide resistance.
Asunto(s)
Aldehído Deshidrogenasa/metabolismo , Isoenzimas/metabolismo , Hígado/enzimología , Mostazas de Fosforamida/metabolismo , Estómago/enzimología , Acetaldehído/metabolismo , Aldehído Deshidrogenasa/aislamiento & purificación , Animales , Cromatografía por Intercambio Iónico , Citosol/enzimología , Femenino , Inactivación Metabólica , Focalización Isoeléctrica , Isoenzimas/aislamiento & purificación , Cinética , Ratones , Ratones Endogámicos DBA , Peso Molecular , Especificidad de Órganos , Mostazas de Fosforamida/síntesis química , Especificidad por SustratoRESUMEN
Bis(2-chloroethyl)phosphoramidic dichloride [MP(O)Cl2, M = N(CH2CH2Cl)2] has been used as the starting material for the synthesis of O-aryl phosphorodiamidates having the general structure MP(O)(NHR)OAr: 9, R = H, Ar = 4-NO2C6H4; 10, R = H, Ar = C6F5; 11, R = C6H5, Ar = C6F5; 12, R = 4-MeC6H4, Ar = C6F5; and 13, R = 4-EtOC6H4, Ar = C6F5. The phosphorodiamidic chloride precursor to 13 (14) was also isolated. Kinetics for the base-catalyzed hydrolysis of compounds 9--13 were investigated by UV and NMR methods and are considered in connection with service of these compounds as pro(phosphorodiamidic acid mustards) [MP(O)(NHR)OAr leads to MP(O)(NHR)OH] via an E1cB mechanism involving the intermediacy of a mustard-bearing metaphosphorodiimide [MP(O)=NR]. Anticancer screening tests against L1210 lymphoid leukemia in mice indicated that 9--14 are inactive; similar negative results were obtained with the KB cell culture, except in the case of 14 which was marginally active.
Asunto(s)
Mostazas de Fosforamida/síntesis química , Animales , Carcinoma de Células Escamosas/tratamiento farmacológico , Femenino , Humanos , Hidrólisis , Técnicas In Vitro , Cinética , Leucemia L1210/tratamiento farmacológico , Masculino , Ratones , Neoplasias de la Boca , Neoplasias Experimentales/tratamiento farmacológico , Mostazas de Fosforamida/uso terapéutico , Relación Estructura-ActividadRESUMEN
The synthesis of the title compound from 3'-amino-3'-deoxyadenosine in 40% yield is reported. 3'-Amino-3'-deoxyadenosine was made by an improved synthesis in 12 steps from inexpensive D-xylose in 15% overall yield. Both isomers of the title compound, separated by column chromatography, possess confirmed activity against KB tumor cell cultures.
Asunto(s)
Nucleótidos de Desoxiadenina/síntesis química , Nucleótidos Cíclicos/síntesis química , Mostazas de Fosforamida/síntesis química , Animales , Carcinoma de Células Escamosas/tratamiento farmacológico , Nucleótidos de Desoxiadenina/farmacología , Nucleótidos de Desoxiadenina/uso terapéutico , Humanos , Técnicas In Vitro , Leucemia L1210/tratamiento farmacológico , Métodos , Ratones , Nucleótidos Cíclicos/farmacología , Nucleótidos Cíclicos/uso terapéutico , Mostazas de Fosforamida/uso terapéuticoRESUMEN
Nine representatives of the title series of compounds [(ClCH2CH2)2NP(O)(NH2)ON = CRR'] were synthesized as potential anticancer prodrugs, based on the possibility of enzymatic reduction of the N-O bond to release the known cytotoxic agent phosphoramide mustard [1, (ClCH2CH2)2NP(O)(NH2)OH]. The dimethyl derivative (2, R = R' = CH3) exhibited a statistically significant, albeit low, level of anti-L1210 activity in mice. Derivative 2, which was shown by 31P NMR measurements to be very stable toward hydrolysis at 37 degrees C over a pH range of 5.7-7.4 (T1/2 congruent to 7-8 weeks), gave colorimetrically detectable amounts of alkylating material upon incubation with mouse liver slices: approximately 3-5% conversion after 20 min at 37 degrees C. A single-crystal X-ray study of 2 revealed an unusual hydrogen-bonded "ladder" and a very similar steric relationship for the NCH2CH2Cl and ON = CCH3 moieties.
Asunto(s)
Antineoplásicos/síntesis química , Oximas/síntesis química , Mostazas de Fosforamida/síntesis química , Animales , Colorimetría , Femenino , Leucemia L1210/tratamiento farmacológico , Hígado/metabolismo , Espectroscopía de Resonancia Magnética , Masculino , Ratones , Oximas/uso terapéutico , Mostazas de Fosforamida/uso terapéutico , Difracción de Rayos XRESUMEN
Phenyl ketone phosphorodiamidates [C6H5C(O)CH2CH2OP(O)NHR1NR2R3] were synthesized in conjunction with an ongoing investigation into the effects of substituents on the dynamical solution chemistry of the metabolites of cyclophosphamide (1a). In contrast to aldophosphamide (3a), which readily interconverts with its cyclic isomer 4-hydroxycyclophosphamide (2a), phenylketophosphamide (14a: R1 = H, R2 = R3 = CH2CH2Cl) exhibited an apparent "resistance" toward an intramolecular addition reaction such that 4-hydroxy-4-phenylcyclophosphamide (13a) could not be detected either spectroscopically (31P or 13C NMR) or chemically (NaCN trapping experiment). Control studies that compared the relative reactivities of 14a and methylketophosphamide [20: CH3C(O)CH2CH2OP(O)NH2N-(CH2CH2Cl)2] revealed that the factors that modulate the ring closure/opening reactions were not peculiar to the phenyl group; however, differences between phenyl and methyl profoundly influenced the rates of fragmentation of 14a and 20. 31P NMR spectroscopy was used to determine the rates at which each compound generated a cytotoxic alkylating agent. Under a standard set of reaction conditions [1 M lutidine buffer with added Me2SO (8:2), pH 7.4, 37 degrees C], the half-lives of 2a/3a, 14a, phenylketoifosfamide (14b: R1 = R2 = CH2CH2Cl, R3 = H), phenylketotrofosfamide (14c: R1 = R2 = R3 = CH2CH2Cl), and 20 were 72, 66, 63, 56, and 173 min, respectively. Analogues 14a and 14b exhibited good anticancer activity against a variety of test systems.
Asunto(s)
Antineoplásicos/síntesis química , Ciclofosfamida/análogos & derivados , Mostazas de Fosforamida/síntesis química , Animales , Antineoplásicos/uso terapéutico , Cianuros , Ciclofosfamida/síntesis química , Ciclofosfamida/uso terapéutico , Semivida , Concentración de Iones de Hidrógeno , Cinética , Leucemia L1210/tratamiento farmacológico , Espectroscopía de Resonancia Magnética , Ratones , Mostazas de Fosforamida/uso terapéuticoRESUMEN
The synthesis of aldophosphamide acetal diacetate and a number of structural analogues is described. These compounds are designed to undergo biotransformation to the corresponding aldehydes in the presence of carboxylate esterases, enzymes that are ubiquitous in mammalian tissue. Several of these aldehydes can theoretically exist in pseudoequilibrium with the 4-hydroxyoxazaphosphorine tautomers; others lack this capability. The half-lives of the acetals in 0.05 M phosphate buffer, pH 7.4, at 37 degrees C ranged from 1 to 2 days. In the presence of 2 unit equiv of porcine liver carboxylate esterase, all of the compounds were hydrolyzed with half-lives of less than 1 min. Although closely structurally related, the compounds exhibited a wide range of cytotoxicities to L1210 murine leukemia cells in vitro.
Asunto(s)
Antineoplásicos/síntesis química , Mostazas de Fosforamida/síntesis química , Profármacos/síntesis química , Animales , Carboxilesterasa , Hidrolasas de Éster Carboxílico/metabolismo , Supervivencia Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Leucemia L1210 , Espectroscopía de Resonancia Magnética , Ratones , Estructura Molecular , Mostazas de Fosforamida/química , Mostazas de Fosforamida/farmacología , Relación Estructura-ActividadRESUMEN
A novel series of cyclophosphamide derivatives of pyrimidine deoxyribonucleosides (6-9) has been synthesized from the corresponding amino nucleosides. Our preliminary findings have shown that three of these cyclophosphamide nucleoside analogues, 6, 7, and 9, have potent inhibitory effects on the replication of L1210 cells in vitro (ED50 = 1.2-1.5 x 10(-5) M). Since cyclophosphamide (cytoxan) has no cytotoxicity under these conditions, our findings indicate that these novel phosphamide derivatives have unusual biological properties which may include a unique mode of activation.
Asunto(s)
Antineoplásicos/síntesis química , Desoxirribonucleósidos/síntesis química , Mostazas de Fosforamida/síntesis química , Nucleósidos de Pirimidina/síntesis química , Animales , División Celular/efectos de los fármacos , Fenómenos Químicos , Química , Desoxirribonucleósidos/farmacología , Leucemia L1210/tratamiento farmacológico , Ratones , Mostazas de Fosforamida/farmacología , Nucleósidos de Pirimidina/farmacologíaRESUMEN
A novel nitroheterocyclic bis(haloethyl)phosphoramidate prodrug linked through lysine to a pteroic acid has been prepared and evaluated as a potential alkylating agent to target tumor cells that overexpress the folate receptor. The prodrug exhibited IC(50) values in the micromolar range and was 10-400-fold less cytotoxic in vitro than the phosphoramidate that lacks the lysine-pteroyl moiety. The data does not support a contribution of the folate receptor to cytotoxicity. In an attempt to determine the basis for the decreased cytotoxicity in the pteroyl-lysyl analogue, compounds were prepared in which the lysine-pteroyl moiety was replaced with lysine alone or with an n-propyl group. The n-propyl and the lysyl analogues were on average 3.8- and 21-fold less potent than the unsubstituted bis(haloethyl)phosphoramidate, respectively. Chemical reduction of the prodrugs followed by (31)P NMR kinetics demonstrated that all of the phosphoramidate anions cyclized to the aziridinium ion at similar rates and gave comparable product distributions, suggesting that changes in chemical activation did not account for the differences in cytotoxicity. It is likely that folate receptor-mediated transport is not sufficient to deliver adequate intracellular concentrations of the cytotoxic phosphoramide mustard.
Asunto(s)
Antineoplásicos Alquilantes/síntesis química , Proteínas Portadoras/metabolismo , Mostazas de Fosforamida/síntesis química , Profármacos/síntesis química , Pterinas/síntesis química , Receptores de Superficie Celular , Antineoplásicos Alquilantes/química , Antineoplásicos Alquilantes/farmacología , Ensayos de Selección de Medicamentos Antitumorales , Receptores de Folato Anclados a GPI , Humanos , Concentración 50 Inhibidora , Cinética , Espectroscopía de Resonancia Magnética , Oxidación-Reducción , Mostazas de Fosforamida/química , Mostazas de Fosforamida/farmacología , Profármacos/química , Profármacos/farmacología , Pterinas/química , Pterinas/farmacología , Relación Estructura-Actividad , Células Tumorales CultivadasRESUMEN
Cyclic and acyclic nitroaryl phosphoramide mustard analogues were activated by E. coli nitroreductase, an enzyme explored in GDEPT. The more active acyclic 4-nitrobenzyl phosphoramide mustard (7) showed 167 500x selective cytotoxicity toward nitroreductase-expressing V79 cells with an IC(50) as low as 0.4 nM. This is about 100x more active and 27x more selective than CB1954 (1). The superior activity was attributed to its better substrate activity (k(cat)/K(m) 19x better than 1) and/or excellent cytotoxicity of phosphoramide mustard released.
Asunto(s)
Antineoplásicos Alquilantes/síntesis química , Activadores de Enzimas/síntesis química , Escherichia coli/enzimología , Nitrocompuestos/síntesis química , Nitrorreductasas/metabolismo , Mostazas de Fosforamida/síntesis química , Profármacos/síntesis química , Animales , Antineoplásicos Alquilantes/farmacología , Línea Celular , Línea Celular Tumoral , Cricetinae , Cricetulus , Ciclofosfamida/análogos & derivados , Ciclofosfamida/síntesis química , Ciclofosfamida/farmacología , Activadores de Enzimas/farmacología , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Humanos , Nitrocompuestos/farmacología , Oxidación-Reducción , Mostazas de Fosforamida/farmacología , Profármacos/farmacología , Relación Estructura-ActividadRESUMEN
A series of 2- and 3-substituted indolequinone phosphoramidate prodrugs targeted to DT-diaphorase (DTD) have been synthesized and evaluated. These compounds are designed to undergo activation via quinone reduction by DTD followed by expulsion of the phosphoramide mustard substituent from the hydroquinone. Chemical reduction of the phosphoramidate prodrugs led to rapid expulsion of the corresponding phosphoramidate anions in both series of compounds. Compounds substituted at the 2-position are excellent substrates for human DTD (k(cat)/K(M) = (2-5) x 10(6) M(-1) s(-1)); however, compounds substituted at the 3-position are potent inhibitors of the target enzyme. Both series of compounds are toxic in HT-29 and BE human colon cancer cell lines in a clonogenic assay. There was a correlation found between cytotoxicity and DTD activity for the 2-series of phosphoramidates; however, there was no correlation between cytotoxicity and DTD activity in the 3-series of compounds. This finding suggests the presence of an alternative mechanism for the activation of these compounds.
Asunto(s)
Antineoplásicos/síntesis química , Hidroquinonas/síntesis química , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Mostazas de Fosforamida/síntesis química , Profármacos/síntesis química , Antineoplásicos/química , Antineoplásicos/farmacología , División Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Hidroquinonas/química , Hidroquinonas/farmacología , NAD(P)H Deshidrogenasa (Quinona)/química , Mostazas de Fosforamida/química , Mostazas de Fosforamida/farmacología , Profármacos/química , Profármacos/farmacología , Relación Estructura-Actividad , Células Tumorales CultivadasRESUMEN
Benzyl phosphoramide mustard (3), 2,4-difluorobenzyl phosphoramide mustard (4), and methyl phosphoramide mustard (5) were examined as lipophilic, chemically stable prodrugs of phosphoramide mustard (2). These phosphorodiamidic esters are designed to undergo biotransformation by hepatic microsomal enzymes to produce 2. The rate of formation of alkylating species, viz., 2, from these prodrugs and their in vitro cytotoxicity toward mouse embryo Balb/c 3T3 cells were comparable to or better than that of cyclophosphamide (1). Preliminary antitumor screening against L1210 leukemia in mice, however, suggests that these prodrugs are devoid of any significant antitumor activity in vivo.
Asunto(s)
Antineoplásicos/síntesis química , Mostazas de Fosforamida/síntesis química , Profármacos/síntesis química , Animales , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapéutico , Fenómenos Químicos , Química , Cromatografía Líquida de Alta Presión , Leucemia L1210/tratamiento farmacológico , Masculino , Ratones , Ratones Endogámicos BALB C , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Mostazas de Fosforamida/farmacocinética , Mostazas de Fosforamida/uso terapéutico , Profármacos/farmacocinética , Profármacos/uso terapéutico , Relación Estructura-ActividadRESUMEN
A series of naphthoquinone and benzimidazolequinone phosphorodiamidates has been synthesized and studied as potential cytotoxic prodrugs activated by DT-diaphorase. Reduction of the quinone moiety in the target compounds was expected to provide a pathway for expulsion of the phosphoramide mustard alkylating agent. All of the compounds synthesized were excellent substrates for purified human DT-diaphorase (k(cat)/K(m) = 3 x 10(7) - 3 x 10(8) M(-1) s(-1)). The naphthoquinones were toxic to both HT-29 and BE human colon cancer cell lines in a clonogenic assay; however, cytotoxicity did not correlate with DT-diaphorase activity in these cell lines. The benzimidazolequinone analogues were 1-2 orders of magnitude less cytotoxic than the naphthoquinone analogues. Chemical reduction of the naphthoquinone led to rapid expulsion of the phosphorodiamidate anion; in contrast, the benzimidazole reduction product was stable. Michael addition of glutathione and other sulfur nucleophiles provides an alternate mechanism for activation of the naphthoquinone phosphorodiamidates, and this mechanism may contribute to the cytotoxicity of these compounds.
Asunto(s)
Antineoplásicos Alquilantes/síntesis química , Bencimidazoles/síntesis química , Inhibidores Enzimáticos/síntesis química , NAD(P)H Deshidrogenasa (Quinona)/antagonistas & inhibidores , Naftoquinonas/síntesis química , Mostazas de Fosforamida/síntesis química , Profármacos/síntesis química , Animales , Antineoplásicos Alquilantes/química , Antineoplásicos Alquilantes/farmacología , Bencimidazoles/química , Bencimidazoles/farmacología , Supervivencia Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Humanos , Cinética , Ratones , Ratones Desnudos , Naftoquinonas/química , Naftoquinonas/farmacología , Trasplante de Neoplasias , Mostazas de Fosforamida/química , Mostazas de Fosforamida/farmacología , Profármacos/química , Profármacos/farmacología , Relación Estructura-ActividadRESUMEN
A series of sulfonyl-group containing analogues of aldophosphamide (Aldo) were synthesized as potential anticancer prodrugs that liberate the cytotoxic phosphoramide mustards (PM, IPM, and tetrakis-PM) via beta-elimination, a nonenzymatic activation mechanism. Kinetic studies demonstrated that all these compounds spontaneously liberate phosphoramide mustards with half-lives in the range of 0.08-15.2 h under model physiological conditions in 0.08 M phosphate buffer at pH 7.4 and 37 degrees C. Analogous to Aldo, the rates of beta-elimination in all compounds was enhanced in reconstituted human plasma under same conditions. The compounds were more potent than the corresponding phosphoramide mustards against V-79 Chinese hamster lung fibroblasts in vitro (IC(50) = 1.8-69.1 microM). Several compounds showed excellent in vivo antitumor activity in CD2F1 mice against both P388/0 (Wild) and P388/CPA (CP-resistant) tumor cell lines.