IE39282B1 - Radionuclide labelled phosphonic acids for the treatment of calcific tumors - Google Patents

Abstract

1453667 Anti-tumour compositions PROCTER & GAMBLE CO 22 May 1974 [23 May 1973] 22847/74 Heading A5B [Also in Division C2] Calcific tumours in animals are treated by administering to the animal a safe but effective amount of a phosphonate, tagged with a <SP>32</SP>p or <SP>33</SP>P radionucleide, selected from compounds of the formulae:- where each R is H or CH 2 OH and n is 3 to 10; where R 1 is H, C 1-20 alkyl, C 2-20 alkenyl, aryl, phenylethenyl, benzyl, halogen, OH, NH 2 , substituted amino, -CH 2 COOH, -CH 2 PO 3 H 2 , -CH(PO 3 H 2 )(OH) or -[CH 2 C(PO 3 H 2 ] n -H, where n is 1 to 15,. and R 2 is H, alkyl, NH 2 , benzyl, halogen, OH, -CH 2 COOH,-CH 2 PO 3 H 2 or -CH 2 CH 2 PO 3 H 2 ; where n is 3 to 9; where each R 3 is H or alkyl; where n is 2 to 4; and (X) where X and Y are each H or OH; or a pharmaceutically acceptable salt of such a compound. A chemotherapeutic agent, selected from alkylating agents, antimetabolites, antibiotics, vinea alkaloids, hormones, enzymes, hydroxyurea and procarbazine, may be administered after the administration of the tagged compound. Specified chemotherapeutic agents are mechlorethamine hydrochloride, triethylenemelamine, thiotepa triethylenethiophosphoramide, bulsulfan, chlorambucil, cyclophosphamide, melphalan, BCNU, 6-mercaptopurine, DON, azaserine, methotrexate, 5-fluorouracil, cytarabine, actinomycin D, dactinomycin, mithramycin, daunomycin, nitomycin C, vinblastine sulphate and vincristine sulphate. Preferably the dosage level of the tagged compound is 0.2 to 20 millicuries. [GB1453667A]

Description

39293 This invention relates to radiopharmaceuticals and nore particularly to such materials suited for use in treatment of calcific tutors il.e., osteoblastic tumors), particularly primary and metastatic bone tumors and soft tissue calcifying 3 rumors.

Radiation therapy for tumors of various types has been well known for none time. One of the raain difficulties with nil radi.itlon therapy is the lack of specificity of both externally applied radiation and radioactive materials; i.e. in the radiation therefrom Is generalized and has a substantial effect on bone marrow activity and soft tissue which is not tumorous as well as that which is.

Attempts have been made heretofore to provide site selective radiopharmaceuticals with son* success. For example, 15 Storaasll reported In the Journal of the American Medical Association 210, 1077—1078 (1969) that sodium phosphate having incorporated therein the radioactive isotope of phosphorous having an atomic weight of 32 (^P) had a measure of selectivity for osseous tumors. The selectivity achieved by this approach 3028 2 Is apparently related to the high uptake of phosphorus In tumors a? roported by Anihilorl In Experlentln 28 Number 9, lOR6—7 (1972) but Is insufficient to allow an effoctlve dose of radioactivity to be directed to a bone tumor without simultaneously directing a damaging dose to unaffected bono, bone marrow and other soft tissue cells. Consequently radioactive phosphates (and polyphosphates) have been restricted to at* In extreme or terminal bone tumor casos vtiero the high risks of additional damage were warranted.

Prior to the present invention, no compound or method existed whereby effective therapeutic or pain relieving doses of radiation could be directed to bone tumor sites without a substantial risk of radiation damage to the remainder of the body.

A host of mono-, dl- and polyphosphonates have recently been found to exhibit generaMzud attraction to osseous sites and are useful in inhibiting the anomalous deposition and mobilization of calcium phosphate in animal tissue as disclosed In r.S. Patent 3,683,OAQ. It has now been discovered »-h»t certain mono-, di-, and polyphosphonates wherein a radionuclide selected from the group 12P and * *P is used in place of stable *lP exhibit unexpectedly and usefully high selectivity for calcific tumors in addition to the hiqh uJt generalize calcific selectivity formerly associated therewith.

Such "tagged" or radioactive phosphonates have, as will hereinafter be described, sufficient selectivity for primary and aetastatic bene tumors that effective amounts of ^P or ^P radionuclide can be delivered to the tumor site without adversely affecting non-tumorous sites. 39282 Selectivity .in great .is 4o.*l Imm born demonstrated In • logs with proven osteoqenic sarcoma; I.e. the tumorous site can contain 4o times as pinch radioactivity as the correspon-•ling hut opposite and unaffected bone (hereinafter referred to as the controllateral hone) or as adjacent normal bone (as in the sternum or spine area).

The Invention provides a novel radioactive phosphonate 32 33 or polyphosphonate compound tagged with a P or P radionuclide selected from conpounds of the formulas II I TOjHj (I) wherein each R is hyrirooen or CHjOH and n is an integer of from 3 to 10» P»2 Rj—C R2 (XI) Jo3Hj wherein Rj is hydrogen, alkyl containing from 1 to 20 carbon atoms, alkenyl containing from 2 to 20 carbon atoms, aryl (e.g. phenyl or naphthyl), phenylethenyl, benzyl, halogen (e.g. chlorine, bromine or fluorine), hydroxyl, amino, substituted amino (e.g. dimethylamino, dlcthylamlno, N-hydroxy-N-ethylamino or acetylamlno, —CH2COO», CH2POjH2 —CH(P03H2) (OH), —CcH2C(P03llJnH where n is 1 to 15, and R2 is hydrogen, alkyl (e.g. methyl, ethyl, propyl or{hutyl>.amino, bensyl, halogen («.g. chlorine, bromine or fluorine), hydroxy!, —-CHjCOOH, —CH2P03H2 or —CH2CH2POjH2 (tH2>n "I » PO,H 3 2 CH- -OH (III) P03H2 3 1* :: 8 i wherein n is an intencr of from 3 to 9» pHa «3-c-«3 R, H203p— C C-p03H2 (iv) Rj Rj wherein each »3 is hydrogen or Cj—C4 alkyl (e.g. methyl, ethyl, propyl or butyl)t I (=r2)n I c-po3h2 (v) -C-P03H2 wherein n is an integer of from 2 to 4» (vi) at 0 1 hopoh I COOH I coon I (VII) f- I h ?°3»2 IcU- .ind tuns-) isomrrr 0 4- ■on * i i HO—P—0—C—Cll, I 1 6 ho-*>—on I o I H—C I (VIII) CWP—OH I OH C I O-rP—OH j>R oh I II M—OH I I to (ix) c c I i H o—r OH I oh and (X) X- COOH I COOH I -C—V I I P03H2 3«2 - 5 - wherein X and Y .ire e.i«-h hydroqen or hydroxy; and the pharmaceutically .iccept.ible silts of each of the forcgolno .iclds, e.g. alkali metal (e.g. sodium or potassium), alkaline i-irth metal (e.g. calcium or magnesium), non-toxic heavy metal (e.g. stannous or Indium) and ammonium and nv>rv-, di- or triethanolanine salt*.

In Its method aspects, the present invention comprises systemlcally administering safe hut effective amounts of iurh c<jm|tounds to a non-human animal afflicted with a calcific Iunor.

The preferred rad<«».ictlve phosphonates are those containing the radionuclide 'V. fultable polyphosphonates of the above formula C) Include propane-1,2,3-trlphosphonlc acid: butane-1,2,).4-tetr> phosphonic acidj hexine-l,2,3,4,5,6-hexaphosphonic acid; hexanc - 1 - hydroxy - 2,3,4,5,* - pentaphosphonlc acid; hcxanc - 1,6 - dlhydroxy - 2.1,4,5 - tctraphosphonlc acid: pentane - 1,2,3,4,5 - pentaphosphonlc acid; heptane - 1,2,1,4, 5,6,7 - heptaphosphontc acid; octane - 1,2,3,4,5,6,7,8 - ccti-phosphonlc acid, nonane - 1,2,3,4,5,6,7,8,9 - nonaphosphonir acid; decane - 1,2, 3,4,S,#», 7,8,•*, 10 - decaphosphonlc acid; .ind the pharmaceutically acceptable salts of these acids, e.g. sodium, potassium, calcium, magnesium, ammonium, trlethanolammonium, dlethanolamnonlun, and monoethanol-ammonlum salts. l'hosphorus-32 and phosphorus-33 are commercially available. The materials can be reacted conventionally to forr PClj. PC13 Is a suitable starting material for all of the syntheses herein outlined. PCl-j car. be made Into suitable intermediates for the disclosed structures by the following reactions: 3ftasa PClj ♦ SlljO—> HPO^Hj ♦ 3HCI PCI, ♦ 3F0I1—-> P(OR), ♦ 3IIC1 A 2P(0Rj) ♦ liro3M2—> 3HP03R2 Na ♦ Hro3R2—> NaPOjRj ♦ l/2Hj wherein R la an organic radical.

A useful outline of reaction* to form polyphosphonfttea from these aaterials la found In *Toplea In Phosphorus r> Chealstry* (Chapter 7, •Ollgophosphonates*, Miley* 1972).

Propane-1,2,3-tripho»phonlc acid and salts thereof can be prepared by a process disclosed In the U.S. Patent 3.743.<88.

Butane-1,2,3,4-tetraphosphonlc acid and salts thereof lo can be prepared by a process disclosed in the 0.9. Patent 3,755,504.

The hl9her aliphatic vicinal polyphoephonatea and salts thereof can be prepared by the process disclosed In U.S. Patent 3,584,015.

IS toong the suitable polyphosphonates encos^assed by the above formula (II) are ethane-l-hydroxy-l,l-diphosphoi*ic acid; Mthanediphosphonic acid; methanehydroxydlphosphonic acid; eth.ine-l,l,2-triphosphonic acid; propane-1,1,3,3-tetraphosphonlc acid; ethane-2-phenyl-l,l-dlphosphonic acid; JO ethane-2-naphthyl-l,l,l-dlphosphonlc acldi mthanephenyldl-phosphonic acid} ethane-l-amlno-l,l-diphoaphonlc acidj methanedichlorodiphosphonlc acid; nonane-5,5-diphosphonlc acid; n-pentane-l,l-diphosphonic acid; methanedifluorodi-phosphonlc acid; nethanedibromodlphosphonic acid} propane-2,2rdlphosphonic acid; ethane-2-carboxy-l,1-dlphosphonic acid; propane-I-hydroxy-1,1,3-trlphosphonic acid; ethane-2-hydroxy-1,1,2-triphosphonic aeld; ethane-l-hydroxy-1,1,2- :» 0 2 H 2 I r lphosphonlc acid; prop.tn#--1, l-«1lphony 1 -2, 2-dlphosphonlc acl'l; n«>n.ino-l, l-dlphn*phonlc <icld; hrxadecane-1,1-dlphosphonlc «cl«J; l>»»nt-4-ene-l-hydroxy-l, 1-dlphosphonlc sclrtj octadee-9-ene-l-hy«lroxy-l , 1-dlphosphonlc acid; 3-phenyl-l, l-dlphosphono-prop-r> /-»»nei octane-l,l-dlpho*phonlc acid; dodecane-1 * 1-dlphosphonlc .ir*Idi phenylamlnomethanedlphosphonlc acid; naphthy1amino-mrthanedlphosphonic acid; N,N-dlmethylaminomethan*dlphoaphoni': artdi N-(2,2-dlhydraxyethyl)-aralno»ethanedlphosphonlc acid} N-acatylamlnomethanedlphosphonlc acid; amlnomethanedlphosphonlc lo .n idi dihydroxymethanedlphosphonlc acldmnd the pharmaceutical ly acceptable salts of these acids, e.g., sodium, potassium, c.tlcium, magnesium, stannous, indium, ammonium, trlethanol-ammonium, dlethanolaramonlum, and ronoethannlaroonlua salts.

Bth*ne-1-hydroxy-1,1-dlphosphonlc acid, an especially l*> preferred polyphosphonate, has the molecular formula ni^C(Oll) IPOjHj)^. (According to nomenclature by radicals, the acid might also he named 1-hydroxyethylidene dlphosphonlc acid.) While any pharmaceutlcally acceptable salt of ethane-1-hydroxy-1,l-dlphosphonlc acid can be used In the practice of 2(> this invention, the disodlum dlhydrogen salt Is preferred. The other sodium, potassium, ammonium, and mono-, dl-, and trlethanolammonium salts and mixtures thereof are also suitable provided caution is observed In requlatlng the total Intake of cation epeclcs in the salt composition. These compounds can 2r< be prepared by any suitable method, however, an especially preferred method is disclosed In British Patent Specification No. 1,131,916.

Methanehydroxydlphosphonlc acid and related compounds operable herein can be prepared,for example, by reaction of 3n phosgene with an alkali metal dlalkylphoaphlte. \ complete - 8 - :i 9 a 8 a description of these compounds and a method of preparing 5.imc in found in DrJtinh Patont t lcat Ion No. 1, HO,')8u.

MethancdIhydroxyrilphosphonlc acid and u.ilt-i useful heroin .ind a method Tor preparing sane are disclosed In U.S. Patent 5 3,497,313.

Methanedlphosphonic acid and related compounds useful herein are described in detail in U.S. Tatent 3,213,030. A preferred method of preparing such compounds is disclosed in U.S. Patent 3,251,907. 10 Ethane-l,l,2-triphosphonic acid and related compounds which can be used in the compositions of this invention, as well as a method for their preparation, are fully described in U.S. Patent 3,551,339.

Propane-l,l,3,3-tetraphosphonic acid and related com-15 pounds useful herein, and a method for preparing su« are fully disclosed in British Patent Specification No. 1,136.619. The higher methylene-lntcrrupted methylene diphosphonate (i.e. those wherein is —^CH2 • wherein n is 2 to 15 ran be prepared by the polymerization of ethylene-1,1-2o diphosphonate.

Pentane-2,2-diphosphonlc acid and related compounds can be prepared in accordance with the method described by G. M. Kosolopoff in J. Amer. Chem. Soc.. 75, 1SOO (1953).

Suitable phosphonates of formula (HI) above Include the 2f> Col lowing t Methanecyclobutylhydroxydiphosphonic acid riethanecyclopentylhydroxydiphosphonic acid Nethanecyclohexylhydroxydiphosphonlc acid Methanecyclohepty1hydroxydiphosphoni c ac id 30 Methanecyclooctylhydroxydiphosphonic acid - 9 - 3 9::« Methanecyclonony Ihydroxyd * • -tt'isplionii* ."«c t >.1 Methan«rryclodecy Ihydroxydi pt-.ofiphi -ur nc id.

E.irh of the sodium, pot'is91un, calciun, nngiu»sj.um, r.L.innous, Indium, amnoninin, -v>noothanol iro-onl«ifi», di^H-ano!-5 amnonliun jnd trlethrtnolomnonlum salts of the above rcclted mcth.mccycloalkylhydroxydiphosphonlc acids as well any other pharmaceutical^ acceptable salt of these acids, al3o selectivity migrate to the skeletal tissues.

The phosphonates of formula (HI) can be prepared by lo method disclosed In "Topics in Phosphorus Chenlstry", Chapter 7, "oiigophosphonates", Wiley 1972.

The preferred phosphonates of formula fIV) for the purpose of this Invention are tr is (phosphonoir.ethyl) amine j r r i.:; (1 -phosphonoethyl) amine; iris (2-phosphoi>o-2-propy 1) .-win"; IC ,»r.d their phamac'jut lcaliy acceptable salts. Tris(phosphono-irethy) )aminc is especially preferred. The following are ^x^mplary of compounds which can also he used. ( i) his(i»tio3phvu»c4<w-thyI)-1-pf'vjsphoncH-t.hv I u<«ln*i (t>) i>u (i>iiost>h'-n'i0b-t fty 1) -2-! ■>>o,,«j'hon»• -.'-pr o|<y I aw ln~«; .■<t (r) Ms» (1-pJ.osuhon«>ethyl) phosphonvw.ethyl «nlue: (d) Mj»(2-phosphono-2-propyl)phosphonomc>chyl amne; (o) t r i s(I-phosphonc-1-penty1)ami ne: (f) »>ls (ubosphonoptothyl) 2-phosphono-2-hc.xyl ami::e; and (<j) thsi pharmaceutical^ acceptable salts of acids (3) 2r» to (f), e.g., sodiunt, potasslur, calcium, iwgneslun, ammonium, trlethanolamrsonlur., vilethanolasunonlum, and nonooth'inolanononiun salts. The tris(phosphonoalkyl)amines can be prepared, for oxwrifla, t>y firr,t preparing the corrcspi.n'1!no oster In iO x-coj-.i.jnco with the rrcnoral reaction: - 10 - 39382 R1 0 *i 1 r 1 1(»0)2P(0) (ll)4"}C»0>NH3 > C<r0)2P"C J31® 2 *1 wherein R is alkyl and Rj and R2 are hydrogen or lower alkyl.

The free acids can he prepared by hydrolysis of the ttater using strong mineral acids such as hydrochloric acid. 5 The salts are, of course, prepared by neutralising the acid with the base of the desired cation. The preparation of tristphosohonoalkyl)amines is fully disclosed in Canadian Patent 753,207.

The phosphonates of foroula (V) includc the followingt lO (1) 3,3,4,4,5,5-hexafluoro-l,2-diphosphonocyclopent-l-ene; (2) ^,3,4,4-tetraflnoro-l,2-'1i"K'>sphonO'.-yclobut-l-*ne» an-1 (3) i,3,4,4,5,5.6,6-occafluoro-l,i-dlphonphonocyclohex-1 - enc.

The perfluorodlphosphonocycloalkenes can be prepared, for 15 example, by reacting trlalkyl phosphites with 1,2-dlchloroper-fluorocvcloalk-l-enes in accordance with the procedures fully described by Frank in J. Org. Chen., 31. No. 5, p. 1521.

The ohosphonate of formula (VI) is referred to herein as cyclic tetraphosphonlc acid. This compound and Its 20 pharmaceutically acceptable salts can be prepared by any suitable method, however, an especially preferred method is <ll»clos*l In British Patent Specification No. 1,079,340.

The ohosphonate of the above formula (VII) is ethene-1,2-dicarboxy-l-phosphonic acid. Pharaaceutically acceptable 25 salts of this acid, e.g., sodium, potasalun. calcium, magnesium, stannous, indium, Ammonium, triethanolammonium, dl-•sthanola/iimoniun, and monocthanolammonlun 9altn, are a]so suitable herein. While the above formula (VII) is represen-t.lvc cf the cl s-laomer, the corresponding trans-lsoner is - 11 - .{9 28 2 also useful herein. Reference hereinafter to ethene-1,2-dicarboxy-l-phosphonic acid or salts thereof, unless otherwise specified, is Intended as contemplating the els- and trane-iaosars and mixtures thereof. 5 Ethene-l»2-dlcarboxy-l-phosphonic acid can be prepared by reaction of an ester of acetylenedicarboxylic acid and a dialkyl phosphite followed by hydrolysis and saponification. This method is More fully described in U.S. Patent 3,584,124. The phosphonate of the formula VIII can be made in relit arrangement reactions of the typei 0 1 HO P OU N*n II O ONa I ill* C1CH,—-C—-OH NaOU O—P—C—C—P—O ♦ Na.HPO. ♦ NaCl 2 I ——> III 24 BO P OH 25 C NaO H ONa The phosphonate of formula IX can be made by the method of German Offenlegungsschrift 2,026,078.

Carboxyphosphonates of the above formula (X) Include is ethane-1,2-dicarboxy-l,2-dlphosphonic acids ethane-1,2- <Ucarboxy-l,2-dlhydroxy-l,2-diphosphonlc acid» ethane-1,2-clicarboxy-l-nydroxy-l,2-dlphosphonic acid; and the pharmaceu-tlcally acceptable salts of these acids, e.g., sodium, potassium, calcium, magnesium, ammonium, tr lethanolaimonium, dl-2o ethanolammonium and monoethanolanvnonlum salts.

Ethane-l,2-dicarboxy-l,2-diphosphontc acid, a preferred carboxyphosphonate herein, has the molecular formula CH(COOH)CPOjH2)CH(COOH)(POjHj). The most conveniently cry-dtalllzable salts of this acid are obtained when three, four 25 or five of the acid hydrogens are replaced by sodium. - 12 - 3U4ti2 WhtJf nny phariu.icou i t: 1 uy acceptable snlt of othane-1 ,2-dtcarboxy-l,2-dlphosphonic acid can he used In the practice of this invention, the tetrasodiun dthydrogen salt, the trisodium trihydrogen salt, the dlsodium totrahydrogen 5 salt, the monosodium pentahydrogen salt, and the mixtures thereof are preferred. The other sodium, potassium, ammonium, and mono-, di-, and triethanolaanonium salts and mixtures thereof are also suitable.

Kthane-l,2-dlcarboxy-l,2-dlphosphonlc acid and suitable lo salts thereof can be prepared in any convenient manner. For example, the reaction described by Pudovik In "Soviet Research on Organo-Phosphorun Compounds", 1949—1956, Part III, S47, 86c, can be used to prepare the es*-- of ethane-1f 2-dlcarboxy-1,2-dlphosphonlc acid which in turn can, by ordinary hydroly-15 sis reactions, be converted co the free acid form. Neutralisation by alkali compounds such as sodium hydroxide, potassium hylcoxlde or carbonates can be used to prepare n desired salt cf the acid, h more detailed description of the preparation of these compounds is described In U.S. Patent 3,562,166, 2o which also gives a description of the preparation of the compound (VIII).

Kth*ne-1,2-dlcarboxy-l,2-dlhydroxy-l,2-dlphosphonlc acid .»n<l related compound* useful herein can L*o prepared hy reaction of an es».er of ethane-1,2-di';arboxy-l,2-dlpho*i>honir ?r> a.id *nd ar. alkali nn-tal hypohalite followed by hydrolysis and saponification. This method is more fully described in U.S. Patent 3,579,570.

Mixtures cf any of the foregoing phosphonlc acids and/or salts can be used in the practice of this invention. 30 Suitable dosage levels of the radioactive polyphosphonates J 3 - .->f » h«» invr*nti<'fi » y f • i. ■.. 1 i cnripi !:!'> ').* to 2'j mllli- "'urii-a (mCi). The uniq«ir> low nnft-t ir.sMp retention can allow even higher levels. Hreferably the compound (or composition comprising two or more compounds) 13 prepare.d from high activlt" 'VI3 and care is taken to assure that all or a high proporticn of the polyphosphonate be tagged in order to minimize the total do9age of polyphosphonate roquired to ■administer the desired radioactive dose. Also, the phos-(•hnn.ite is preferably used shortly (e.g., within three days) aft».»r preparation. A specific <«ctivity of at least O.Ol nCi/mg of phosphanate at the time of administration Is preferable. Excessive total doses of polyphosphonates.

It is believed,can result In a saturation of the region of the active tumor and an "overflow" of the polyphosphonate »without regard to its ' "vel of tagging) to non-tumerous are-i s.

Doth, for storage and use, it is preferred that the compounds of thij Invention he in dilute, sterile, pyrogen free, .»'}iie«)us i-eiution torn, e.g., about 2 mg/ml. Such ^ nolntion induce.. 'he probability of self degradation of thr* phosph ates due to the radiation given off therefi-or; and is suitable for direct infection to the patient, preferably lntr.v/eneous ly . "uch solutions can he further di Luted if desired. Suitable •}la-;r. ^l.ilr. with rnd'ntion resistant olaj.tirr or rubier sea's are preferably used to pjekage the solutl -ns.

The following Examples are illustrative of the present j riven t ion : "XAMPLK 1 A Standard Poodle with an advanced bone t imor In the leff ulni w,is administered four lnjectioi.s (>ne par d.iy on 30282 successive days) of a 32P-tigyed <11 sodium ethane-l-hydroxy-1,1-fHphosphonate solution in the cephalic vein.

The solution was prepared by a reaction of the type described in the afore -mentioned U.S. Patent 3,400,14** and In .'ACS; Vol. 49, pp. fcll?, et seq. Specifically, PClj 0.80 ail, 4 mM, 17 mCi) and o.io ml of yiacl.ii aco*. ic ucld w->r«> qontly mixed at roon temperature. Thn acetic acid served both as a reactant and • solvent. 0.3 ml of water was then added dropwlse over a three minutes period. The mixture was slowly heated (over two hours) to 14S°C and held at this tenperature for four hours, 8 ol of water was then added to the reaction flask along with a mixing chip Olengar qranule) and the solution wis refluxed for 40 hours at 14S°C. resulting in a product consisting primarily of ethane-l-hydrcxy-l,l-dipK->sphonlc acid. The acid solution was a«ljuntcd to a pit nf about 5 with NaOH resulting in di --odium etSane-1-hydroxv-1 4-dlphos»phonute. Water w»s added t" .idjus* the concentration to 12.9 ml of HjO/mM of phos-phonate ind formula 3A anhydrous ethanol wis added (2.5 parts ethanol/part phospnonate solution) slowly with vigorous solution to precipitate the diphosphonate. The precipitate was vacuum filtered, via shed with eth.-\nol and ethyl ether and dried. The dry product was stored ir. aqueous solution (SO mg/ml) in order to minimize radiation decomposition.

About 0.3 mCi per injection was used for a total of 1.2 mCi. A noticeable reduction in pain (as evidenced by. Inter alia, increased activity) resulted. There were no ncMcrablr signs of radiation sickness, because oT the advanced state of the turgor yhen therapy corraenred and the low dosages of radioactivity administered therapy was incomplete and anputa-tion was ultimately indicated. 3 9 2 8 3 EXAMPLE 2 A Great Pane with an advanced osteogenic sarcoma In the left radius was treated as In Example 1 out with five dopes of O.S mCi each on successive days. About 10 days 5 after the final administration of the radioactive drug, pi I n reduction and Increased activity was noticed. No radiation sickness wan noticed. Again, the tumor was in such an advanced stage when the dog became available for therapy that the therapy was only partially succcnsful aiwl lO <>uU.anacla was ultimately indicated. A post-mortem was conducted; the tumorous bone was found to have 40 times (on a mCi/gm basis) as much radioactivity as the contralateral bone. In addition, gross pathological observation suggested that lysis (dissolution of the tumor) had occurred to some IS extent.

EXAMPLE 3 A Vizsla with a bone tuanr in the right proxinal humerus was treated as in Example 1 except that about 1 mCi was administered on each of four successive days. About four JO days after the last injection the dog progressed to the point of running using the leg which it previous1/ had "carried".

EXAMPLE 4 A 7<> kilogram adult with a bon»* tumor is administered intravenously dlsodiun-ethane~l-hydroxy-l,l-diphosohonate hav-25 ing *2P incorporated therein (8o mCi/gri) in five doses of 2 nC1 each on successive days. About 4 days after the last injection, a noticeable reduction In bone pain is observed.

Oichloromethanediphosphonic acid; tris(phosphonomethyl)-amine; methane cyclohexylhydroxydlphosphonlc acid; 3,3,4,4,5,5-30 hexafluoro - 1,2 - diphosphonocvclo - pent - 1 - ene; cyclic - 16 - 3&aea tetraphosphonlc acid; ethane - 1,2 - dicarboxy - 1 - phosphonlc •icldj and ethane - 1,2 - dicarboxy - 1,2 -dlphosphonlc acid l».«vin<j Incorporated therein are substituted for the ethane-I-hydroxy-I, l-<liphoH phonic acid and simular results are obtained. * The radio active phosphonates herein enumerated also are cutted for combined therapy with chenotherapeutic drugs. Such drugs can be conveniently classified in the following groups; alkylating agents* antimetabolites, antibiotics, vinca alkaloids, hormones, enzymes (e.«., 1-asparginase) hydroxyurea and procarba-10 zlne (N-isopropyl-a-(2-methylhydrazino)-p-toluaalde nonohydro-i-hlorlde). In conblned therapy using radioactive phosphonates and chenotherapeutlc agents the polyphosphonate will t<e admin!s-tcre<l first and the other chenotherapeutlc agent will be administered In conventional doses (i.e., doses on the order of those used for such chcmothe-apeutle agents alone) beginning at from 1 to 20 days after the administration of the radioactive phosphonates. This approach takns a«l vantage of the fact that »-h«raothcrap<*utlc agents work durlnq the active cycle of rolls r^, •int' that radioactivity can stimulate transit to 2r. of cells fr^ra tho testing (Gn) state int" the active cell cycle.

Alkylating agents suitable for use in combined therapy with radioactive phosphonates include nechloroeth-vnlnc hydrochloride, <2,2,-di'*hloro-N-n»e»thyldiethylainine hydrochloride), t.ri<*thylenemelaraine (2,4,6 - tris(L - azirldinyl) - s - triazinc), 2r> Ltiiotopa (triethylenethiophosphoroamide), busulfan (1,4 - bls-(nethanesultonoxy)butanei, chlorambucil (4 - p - Cbls(2-rhloroethyl)ae»in<iJphenyl butyric acid), cyclophosphamide (1-bis{2 - chloromethyl)amlno - 1 - oxy - 2 - az» - 5 - oxaphos-phorldin), melphalan(p - (di - 2 - chloroethylamlno)phenylalenIns) 3n and BCNU (1,3 - bls(2 - chloroethyl) - I - nitrosourea. - 17 - J a 54 8 z Ant imoJ..ilx>l iten it ii»lr Uu use fn foiiibln»««l tl«*r*py with radioactive phosphonates indudc »>-picr.capfcop»iri nc, (6-|.urinot'ii"l), OON (#» - diazn - *> - oxo - I. - nor - lt-m-ino) , axasorjne (0 - aiazoacoty 1 - L - s«rim ), methotrexate (4-S Amino - v|f> - methylpteroylglutamic acid), S-fluorouracil, cytarabinc II - ft - D - arablnofuranosylcytoslre). Suitable antibiotics include actinonycin D, dactlnomycln, mithranycin, daunotnycin and nitooycin D. Suitable vinca alkaloids include vinblastine sulfate and vincristine sulfate. The above l«> t-hrrentherapeutlc agents and typical dnsnqes t.hercfr<re are 'Inscribed more fully tn the Encyclopedia of Chemical Technology, 2nd Ed. (Interscience, 1971).

EXAMPLE 5 A 70 kilogram adult with a bone tumor is 'Administered I r. i nt ravenously d1sodium-ethane-1-hydro*/-I, T -«ftphosphonattf ii.i'Mng ^^1* incorporated therein (HO mCl/«jm) in flvo <lr>scs ni .» r*'t t»j«;h *>n successive day*. Five days nfter the final in j.-.;tion, administration of 5-fluorouracil at 15 mg/kg/day for five days is be<;un. An increased effectiveness over results 20 produced with either treatment alone results and essentially no side effects beyond those normally associated with the 5-fluorouracil are observed.

EXAMPLES 6—10 *fhen in Examples 1—5 the radionuclide ^P is used instead 25 of ^P substantially the same or similar results are obtained.

Similar results are obtained by adr.inisteriii? other r-neriot hei ap«*ut ic a.jents it. theit -vv.vcrt ion* I do.sages foll'-iwln'j tin. administration of t:»o ra-lioacti ve pf.us;>hon'«t';s !■>> (>no to •is n.mv a:; 29 days thereafter. - 18 - 3838.J It i ippiront trt.it i rosont invention provides a whole new group ot' compounds and nuthods for the treatment "f calcific- tumors. ""ho foregoing ex.ir-plcs .ire illustrative • >n ty. - 19 -

Claims (17)

:»o a H :! C l. A IMS

1. A radioactive phosptit>nat« or polyphosphonate compound faaged with a ^*P or ^3P radionuclide selected from compounds of the formulas It I IojHj (I) wherein each R is hydrogen or CH^OII and n is an integer of from 3 to 10; f°3»2 -C 1 I po3H2 (II) wherein Rj is hydrogen, alkyl containing froa 1 to 20 carbon /ttoma, alkenyl containing fron 2 to 20 carbon atoon, aryl, ;>h«nylethenyl, benzyl, halogen, hydroxyl, amino, substituted amino, —CHjCOOH, —CH2P03H2 —CH(P03H2) (OH) , T — bH2C(P03H2)2]n-H •vher<» n is 1 to 15, and R2 is hydrogen, alkyl, .urine, !»t.-n*yl, h.ilogen, hydroxyl, —CHjCOOH, —ClIjP'^Hj, or -CH2OI-,P<>.jH2j r Wn "I I CH—C I 1 P0jH2 -OH (III) P03M, •.herein n is an integer of from 3 to 9; - 20 - J92B2 ■ 'OH, I 1 - l3 \ / ' \ / R3 11,0,1' C C— PO,H-, (IV) Z 3 | | 32 ^ »3 whp.oin ench Rj i.i hyilroyen or Cj-O^ ilkylj I ;-»3»2 L <cr2)n I (V) -c-po3h2 therein n Is an inteqor of from 2 to 4; n i HOIOH 0 I I (vi) cu.—c—o—p—on I ' COOH COOH O HO-P—OH I I « (VII) r.-■ ■ ^ Icls and trans-) 0 Isomers II o (viri) h—c c I I O—P—OH 0—J>—OH I I OH Oil OH I II »i—P 'Ml I (IX) >: -C and COOH CTHI I 1 1 1 (x) x—e ■:—y II "»l I I I PO,H, PO.,11.. OH 3 2 3 2 wtirtjin X and v arc e.ic'i hydrogen or hyc?roxy; and ♦h-.- :e«»tically accept able salts o' each of the fcrogoing -»Civls.

2. A corpnund according to cl iii" 1 selected from ethane-1-hydroxy-1,1-dtphosphonic acid and pharmaceutlcally acceptable salt3 thereof. - 21 - 9282

3. A compound according t.j ■•iaim 1 selected from di-loromethanediphosphonir acid an<! ph.irr.aceut ical ly acceptable salts thereof.

4. A compound according to claim I selected from tris-(phpaphcnrmethyl)amine and pharmaceutlcally acceptable salts 'hereof.

5. A conpound according to claim 1 selected from methane-c/clohexylhydroxydlphosphonic acid and pharmaceutlcally acceptable salts thereof.

6. A coapound according to claim I selected from 3,3,4,4,5,5-hexafluoro-1,2-diphosphonocyclopent-l-ene and pharmaceutlcally acceptable salts thereof.

7. A coapound according to claim 1 so!«ct*d from the *:*/•-! ir totraphosphonic acid of the formula (VI) and pharnMceur.ically acceptable salts thereof.

3. A coepound according to claim 1 selected fron ethene-1,2-dicarboxy-l-phosphonic acid and pharmaceutlcally acceptable salts thereof.

9. A compound according to claim 1 selected from ethane-1,2-dic^.rboxy-l,2-dlphosphonlc acivl and pharmaceuticals acceptrfhle salts thereof.

10. a dosaqe unit comprising fron 0.2 to 20 milliCurles of a compound according to any of claims 1 to 9.

11. A method for the treatment of calcific tumors in animals comprising systcmically administering to the anLmal a safe but effective amount of a compound according to any of claims 1 to 9.

12. A mothod for 'h<j treatment ot wl'it'ic t'jaors ir. an animal, comprising systcmically administering to rhe anlm»J a tiafc but eff»ctive amount of a compound according to an/ of - 22 - i " ' rJ I'l :» b a M a * * 1. • I . I ' r> '* .t.vi >.>il s.( I" ,1'U.i in i r; t .• r » n<i .1 oh^PioHior- i[>eui 1.: ci>|i nt :s**l "rt • fi : t oi-i alkylating ant 11 rs .in' IhciMT.; viiii' 1 .I I k>t lm t!s ; hormones; en/.yrn's ? >>yd roxyuro." Mid pro'" irtiiizino.

A method for thn treatment of calcific tumors in non-human animals substantially an herein described tn any one of Kxamplos 1 to 3.

14. A compound according to clair. 1 which Is tagged with the ^2P radionucl Ide.

15. A domqc unit. ,*rrrrortfing ».o rlain lo in which the (•'j*tp«>und I:» according to clain J4.

I *>. A mothrid .»(•<*!iftj 1 nq to '*lain 11 i:. wk: irh tli«i compound ij .iTcordlnq to clairr 11.

17. A net not! according to claim 1^ 111 win ?h tfH> (-rvipound is .v •• >rding t'» "lain )4. I . P. K^LI.Y * CO. AGK:rrr» *or the applicants.