ITMI992197A1 - 10-FORMILTETRAIDROFOLATO DEIDROGENASI - Google Patents

10-FORMILTETRAIDROFOLATO DEIDROGENASI Download PDF

Info

Publication number
ITMI992197A1
ITMI992197A1 IT1999MI002197A ITMI992197A ITMI992197A1 IT MI992197 A1 ITMI992197 A1 IT MI992197A1 IT 1999MI002197 A IT1999MI002197 A IT 1999MI002197A IT MI992197 A ITMI992197 A IT MI992197A IT MI992197 A1 ITMI992197 A1 IT MI992197A1
Authority
IT
Italy
Prior art keywords
enzyme
treatment
liver
doses
deae
Prior art date
Application number
IT1999MI002197A
Other languages
Italian (it)
Inventor
Alberto Bartorelli
Original Assignee
Pharmaproducts Uk Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Pharmaproducts Uk Ltd filed Critical Pharmaproducts Uk Ltd
Publication of ITMI992197A0 publication Critical patent/ITMI992197A0/en
Priority to IT1999MI002197 priority Critical patent/IT1314197B1/en
Priority to JP2001501245A priority patent/JP2003501398A/en
Priority to PCT/EP2000/005066 priority patent/WO2000074711A2/en
Priority to DE60004819T priority patent/DE60004819T2/en
Priority to AU59695/00A priority patent/AU5969500A/en
Priority to EP00945698A priority patent/EP1181046B1/en
Priority to ES00945698T priority patent/ES2204649T3/en
Priority to AT00945698T priority patent/ATE247976T1/en
Publication of ITMI992197A1 publication Critical patent/ITMI992197A1/en
Application granted granted Critical
Publication of IT1314197B1 publication Critical patent/IT1314197B1/en

Links

Description

Descrizione dell'invenzione industriale avente per titolo: Description of the industrial invention entitled:

" 1 O-FORMILTETRAIDROFOL ATO DEIDROGENASI COME AGENTE TERAPEUTICO” "1 O-FORMYLTETRAIDROFOL ATO DEHYDROGENASE AS A THERAPEUTIC AGENT"

La presente invenzione riguarda l'uso di 10-formiltetraidrofolato deidrogenasi come agente terapeutico, in particolare come agente citotossico e antitumorale. The present invention relates to the use of 10-formyltetrahydrofolate dehydrogenase as a therapeutic agent, in particular as a cytotoxic and antitumor agent.

La 10-formiltetraidrofolato deidrogenasi è un enzima, presente nel fegato e nel sistema nervoso di mammiferi, di cui non è stato finora descritto alcun uso terapeutico. 10-formyltetrahydrofolate dehydrogenase is an enzyme, present in the liver and nervous system of mammals, of which no therapeutic use has so far been described.

Il cDNA di 10-formiltetraidrofolato deidrogenasi di ratto è stato descritto in J. Biol. Chem. 266(8), 4965-4973, 1991, mentre più recentemente è stato descritto il cDNA dello stesso enzima umano (Biochem. Mol. Biol. Int., 47(3), 407-415, 1999). Rat 10-formyltetrahydrofolate dehydrogenase cDNA was described in J. Biol. Chem. 266 (8), 4965-4973, 1991, while more recently the cDNA of the same human enzyme has been described (Biochem. Mol. Biol. Int., 47 (3), 407-415, 1999).

Metodi per la preparazione deU'enzima ricombinante sono inoltre noti da Protein Expression Purif. 6, 457-64, 1995 e Biochem. J. 306(3), 651-5, 1995. Methods for the preparation of the recombinant enzyme are also known from Protein Expression Purif. 6, 457-64, 1995 and Biochem. J. 306 (3), 651-5, 1995.

Si è ora trovato che la 10-formiltetraidrofolato deidrogenasi di mammifero è in grado di indurre, se somministrata a pazienti o animali portatori di tumori, una marcata risposta citotossica nei confronti di cellule tumorali. It has now been found that mammalian 10-formyltetrahydrofolate dehydrogenase is capable of inducing, if administered to patients or animals carrying tumors, a marked cytotoxic response towards tumor cells.

Tale citotossicità sembra mediata da anticorpi citotossici nei confronti di cellule di tumori umani, in particolare di carcinomi e adenocarcinomi. This cytotoxicity appears to be mediated by cytotoxic antibodies against human tumor cells, particularly carcinomas and adenocarcinomas.

La citotossicità è qualificabile in vitro su cellule HT-29, Kato III e Cytotoxicity is qualifiable in vitro on HT-29, Kato III e

Claims (1)

HepG2 utilizzando metodi convenzionali, basati ad esempio all'uso di kit commerciali quali il kit CDCpK (Pharmaproduct). In particolare, si notava la comparsa di citotossicità nel siero di conigli già dopo un primo trattamento con l'enzima (1 mg/animale in soluzione fisiologica) su cellule Jurkat e Kato III. L’invenzione riguarda pertanto anche composizioni farmaceutiche contenenti come ingrediente attivo una dose efficace di 10-formiltetraidrofolato deidrogenasi. Le composizioni dell'invenzione potranno essere somministrate a pazienti affetti da tumori utilizzando le vie di somministrazione convenzionali per proteine e polipeptidi, ad esempio la via sottocutanea o intramuscolare. Il trattamento può essere ripetuto ed è preferibile un trattamento che prevede somministrazioni distanziate di una-due settimane di dosi comprese tra 0,1 e 20 mg di enzima. Si è inoltre sorprendentemente trovato che è possibile indurre elevata citotossicità anche somministrando l'enzima a dosaggi molto bassi, dell'ordine di 1 , 10<4 >- 1,10<10 >g, per via sublinguale, sotto forma di granuli o gocce di soluzioni o sospensioni idroalcoliche all' 1 % di etanolo, con concentrazioni di principio attivo variabili da IO<"6 >a 10 <10 >M. La 10-formiltetraidrofolato deidrogenasi può essere preparato con metodi convenzionali di DNA ricombinante oppure può essere estratta da fegato di animali, ad esempio da fegato di bovini, ovini o suini. Il fegato di capra si è dimostrato in particolare una fonte abbondante di questo enzima. Il procedimento di estrazione prevede il trattamento dei fegati con soluzioni tamponate a pH 7.4 (PBS) seguito da precipitazione con polietilenglicol 6000 al 15%, cromatografia su TSK-DEAE o DEAE-Sephacell HepG2 using conventional methods, based for example on the use of commercial kits such as the CDCpK (Pharmaproduct) kit. In particular, the appearance of cytotoxicity in rabbit serum was noted already after a first treatment with the enzyme (1 mg / animal in physiological solution) on Jurkat and Kato III cells. The invention therefore also relates to pharmaceutical compositions containing as an active ingredient an effective dose of 10-formyltetrahydrofolate dehydrogenase. The compositions of the invention can be administered to patients affected by tumors using the conventional routes of administration for proteins and polypeptides, for example the subcutaneous or intramuscular route. The treatment can be repeated and a treatment involving one to two weeks spaced doses of doses ranging from 0.1 to 20 mg of enzyme is preferable. It has also surprisingly been found that it is possible to induce high cytotoxicity even by administering the enzyme at very low doses, of the order of 1, 10 <4> - 1.10 <10> g, sublingually, in the form of granules or drops. 1% ethanol hydroalcoholic solutions or suspensions, with concentrations of active principle ranging from 10 <"6> to 10 <10> M. 10-formyltetrahydrofolate dehydrogenase can be prepared by conventional recombinant DNA methods or it can be extracted from animal liver, for example from cattle, sheep or pig liver. Goat liver has been shown to be an abundant source of this enzyme in particular. The extraction process involves the treatment of livers with buffered solutions at pH 7.4 (PBS) followed by precipitation with 15% polyethylene glycol 6000, chromatography on TSK-DEAE or DEAE-Sephacell
IT1999MI002197 1999-06-03 1999-10-20 10-Formyltetrahydrofolate dehydrogenase, useful as a therapeutic agent, particularly as an antitumor agent possessing high cytotoxicity IT1314197B1 (en)

Priority Applications (8)

Application Number Priority Date Filing Date Title
IT1999MI002197 IT1314197B1 (en) 1999-10-20 1999-10-20 10-Formyltetrahydrofolate dehydrogenase, useful as a therapeutic agent, particularly as an antitumor agent possessing high cytotoxicity
AU59695/00A AU5969500A (en) 1999-06-03 2000-06-02 10-formyltetrahydrofolate dehydrogenase as therapeutical agent
PCT/EP2000/005066 WO2000074711A2 (en) 1999-06-03 2000-06-02 10-formyltetrahydrofolate dehydrogenase as therapeutical agent
DE60004819T DE60004819T2 (en) 1999-06-03 2000-06-02 10-FORMYLTETRAHYDROFOLATE DEHYDROGENASE AS A THERAPEUTIC AGENT
JP2001501245A JP2003501398A (en) 1999-06-03 2000-06-02 10-formyltetrahydrofolate dehydrogenase as therapeutic agent
EP00945698A EP1181046B1 (en) 1999-06-03 2000-06-02 10-formyltetrahydrofolate dehydrogenase as therapeutical agent
ES00945698T ES2204649T3 (en) 1999-06-03 2000-06-02 `10-FORMILTETRAHYDROPHOLATE DEHYDROGENASE AS A THERAPEUTIC AGENT.
AT00945698T ATE247976T1 (en) 1999-06-03 2000-06-02 10-FORMYLTETRAHYDROFOLATE DEHYDROGENASE AS A THERAPEUTIC AGENT

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
IT1999MI002197 IT1314197B1 (en) 1999-10-20 1999-10-20 10-Formyltetrahydrofolate dehydrogenase, useful as a therapeutic agent, particularly as an antitumor agent possessing high cytotoxicity

Publications (3)

Publication Number Publication Date
ITMI992197A0 ITMI992197A0 (en) 1999-10-20
ITMI992197A1 true ITMI992197A1 (en) 2001-04-20
IT1314197B1 IT1314197B1 (en) 2002-12-06

Family

ID=11383818

Family Applications (1)

Application Number Title Priority Date Filing Date
IT1999MI002197 IT1314197B1 (en) 1999-06-03 1999-10-20 10-Formyltetrahydrofolate dehydrogenase, useful as a therapeutic agent, particularly as an antitumor agent possessing high cytotoxicity

Country Status (1)

Country Link
IT (1) IT1314197B1 (en)

Also Published As

Publication number Publication date
ITMI992197A0 (en) 1999-10-20
IT1314197B1 (en) 2002-12-06

Similar Documents

Publication Publication Date Title
EP1958643B1 (en) Stable formulations containing enhancing proportions of gamma- and alpha-interferons
NO20053424L (en) Improved pharmaceutical botulinum toxin preparations.
CN109312313A (en) For treating the mRNA therapy of ornithine transcarbamylase deficiency disease
JPH0770195A (en) Sugar-modified interferon
BR0009043A (en) Compound, pharmaceutical composition, and method of treating a disease in an animal in which the activity of the cysteine protease contributes to the pathology and / or symptomatology of the disease
CA2376894C (en) Pharmaceutical composition comprising a protein and an ectoine
ES2322234T3 (en) MODIFIED CITOCINES FOR USE IN CANCER THERAPY.
Guo et al. Head-to-tail macrocyclization of albumin-binding domain fused interferon alpha improves the stability, activity, tumor penetration, and pharmacology
EP1684681B1 (en) A mixture for transdermal delivery of low and high molecular weight compounds
US20200268666A1 (en) Polynucleotides encoding coagulation factor viii
EP1173476A4 (en) Soybean protein nutraceuticals
ITMI992197A1 (en) 10-FORMILTETRAIDROFOLATO DEIDROGENASI
WO1999002120A3 (en) Compositions and methods for reversibly increasing permeability of biomembranes
ES2326530T3 (en) MODIFIED CITOCINES FOR USE IN CANCER THERAPY.
US20220323542A1 (en) TREATMENT OF CYSTIC FIBROSIS BY DELIVERY OF NEBULIZED mRNA ENCODING CFTR
ES2204649T3 (en) `10-FORMILTETRAHYDROPHOLATE DEHYDROGENASE AS A THERAPEUTIC AGENT.
CN115702159A (en) NGR conjugates and uses thereof
On et al. Pathways for drug delivery to the central nervous system
EA026453B1 (en) Use of oxidised avidin for inhalation
JP3522798B2 (en) Method for producing sugar-modified protein
US20230158100A1 (en) Treatment of panx1 associates diseases
JPH01246226A (en) Stable composition containing modified asparaginase
WO2024078729A1 (en) Placenta expressed proteins for use in the treatment of tendon injury
JP2000109434A (en) Inhibition of cell growth and cell growth inhibitor
ITMI982634A1 (en) PHARMACEUTICAL COMPOSITIONS BASED ON PROTEINS.