CA2034242A1 - Macrocyclic tetraaza compounds containing a six-membered ring, processes for their production, and pharmaceutical agents containing same - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE

Macrocyclic tetraaza compounds, containing a six-membered ring, of general Formula I

(I).
wherein is a single or double bond, Q is a nitrogen atom or the residue NH, X1 is a hydrogen atom, a -(CH2)n-R1 group or a

Description

Z0;~4ZL1 2 MACROC~LIC TE$RAAZA COMPOUND~ ~ON~AINING A
SIX-MEMBERED RING, PROCESSES FOR TMEIR PRo~UC~ION, AND PHARMA~EUTICA~ A~ENTS CONTAINING 5AME

Summarv of the Inven.~iQn The invention relates to macrocyalic tetraaza complexing compounds, complexes, and complex salts containing a six-membered rin~. The invent~on furth~r relat~s to agents contalning these compourlds, th~ir u~e as dlagnostic alds and therapeutic agents, as woll as prace~ses for the preparation of these compounds and agents.
Metallic complexes have been scrutinized a6 ea~ly as the beginning of the fifties as contrast media for radiology. The compounds then employed were, however, o~ such toxicity ~hat utilization on human patients could not be considered. It was, therefore, entir~ly surpri~ing to Pind that certain complex salt~
~xhibited adequate.comp~tL~ility for routine administration to human patients for diagnostic purposes.
The first recorded representative of this class of compounds i~ the di~eglumine salt of Gd DTPA ~MAGNEVIST~, .
Schering A.G.; gadoliniu~ ~III) complex o~ -d~ethylenetri~minepentaac~tic acid] descri~ed in Europ~an ~, Pat~nt Applica~ioh Publioation No. 715~4 as a ~ontrast medium for nuclea~ magnetic r~3sonan~:e ima~ing or MRI.
Presently, the compound ls primar~ ly administer~d in c~se ~f di ~eases o~ the central nervous system.
An essen~ial reason ~or the satisfactory ~linical applicabili~y of ~d DTPA resides in its high efficacy in nucleAr magne~iG resonana~ imaging, parti~ulàrly in oonnection with many brain tumors. On account o~

:,~

- 2 - ~0;~4Z~ 2 it.s hi~ll effec:tiveness, Gd DTPA, with 0.1 nunol/kg of body weigh~,_can be util~zed at ~ very much lower dosage than, fol: example, X-ray con'ra~t media, in n~any X-ray examinatlons.
~nother rep~esentative of the complex salts that~ has prover- i.tself well ~or diagnostic purposes is the rnegluntine sal~ o~ Gd DOTA described in German Patent Applicatiorl 3,401,052 Igadolinium(~ complex of 1,4,7,1~-~etraazacyclododecanetetraaceti.c acidl.
llowever, it is desirable to be ahle to tlse ~helates al~o at a higher dosage. This i9 ~he oase, in par~icular, for the detection of cer~ain diseases o~t~ide o the central nervous system Witll the ald of ma~netic resonance i~aging ~MR dia~no~tics), but qui~e e~pecially in case o~ utlli~t~ion of chelates as X-ray contrast media, In order to keep thte volu~ne load on ~he body a~ a m~nimu~, it i~ pre~erabIe to employ highly ~oncentrated chelate 801ution8. ~he ahelates known thus ~ar ~how little ~uitability for this purpose, ~bove all on accourtt of ~heir high osmol~lity.
Therefore, there is a need for ~helates exhibit-ing an osmolality which is lower than ~hat of the previously known ~helates. Ilowever, at the same tinte, 2~ the prerequisites must be met for the use of the~e cot~tpounds on humall patients, as r~gards the 5pacing between the effective do~e and the toxio do~e ~n ani~al tes~ (the ~he~peutl~ ranqe), organ -t~pe~ificity, stability, ~orttrast-enhancing efect, ~ompatibili~y~
as well as sol~tbility of the complex compounds.

~ 3 ~ 20~42~2 Accordir~gly, an objoat o~ th~ invention i8 to make ava~la~e xu~h compounds and agent~, and al~o to provide a ~axim~lly ~imple method for thei~ production.
Thls object is ~et by the p~esent lnvention.
Upon ~urthe~ ~tudy of the ~pecification and appended claim~, furt~er o~ects and advantage~ of this invention will become a~parent to those ~illed in ~he art.
~he complex compounds o~ ~his inven~ion and the solutions prepa~e~ ther~from fUlfill the aforementioned requ~re~nts ln a ~urpr~lng way. ~he~ posses~ reduced osmolality, ~s ~ell a~ a more favorable ~herapeutlç range and/o~ stability an~ shel~ of the chemical ingredients of the solution and/or organ sp~cificity and/or contr~st-enhancing efrect (¢.g., relaxiv~ty) ~ -and~or ~ompatib~lit~ (for ~xample~ reduced oardiov~sc~lar or allergy-type side eff~at~) than th~ previou~ly customary diaqno~tic aid~.
Even without specific measures, the pharm~co-kineti~s o~ the inventive co~pounds per~it. ~n imp~ovement in ~h~ diaghosi3 o~ numerous dioeases. The complexos, for ~he most pa~t, are excre~ed again unchanged and rapidly, ~ that no damaging effec~s are o~erved in spite of a high dose, particularly when using e~en ~ela~ively t~xi~ metallic ion~
~he practical u~age of ~h~ novel complexe~ and compl~xing compound~ i9 al~o facillta~ed by the~r favorable chemioal st~illty ~nother es~ential advantage of the describod ~omplexes ~nd ao~plexing aompound~ is th~i~ extraordinary chemical versatility. The properties can be adapted, not only ~y selec~ion o~ the central atom, ~ut also by the ~hoice o~ variegated sub6tituent~ in the m~crocy~le ~nd~or ~y the ~election o~ ~he salt-forminq compound3, ~o the re~uirement~ regarding ef~ic~cy, p~rmacokineti~
compatibility, ~olu4ility, handling ~bili~y, etc. Thus, it i8 possible ~or ex~mple, to achieve a sp~c~ficity o~
the ~ompound~, very much dssir~blQ in :. :

2034Z~tZ
cl.ia-311c)sti(s al~-l tllerclpy, for structures in the organism, for certaill ~i.ochelllical substances, for metabolic pro-ccsses, for conditions of the tissues or body fluids.
The macrocyclic compounds according to this 5 illVelltiOIl are characterized by general Formula I:
x ZOOC -Cll--N N--Cll-COO~ ( I ), n~
Cll -x ~ooz wherein --- is a single or double bond, Q is a nitrogen atom or the residue NH, 10 Xl is a hydrogen atom, a -(CH2)n-Rl group or a -~CH2~m-(fH)n-cH2OH group OH
wherein n means the numbers 1 to 5, m means the numbers 0 to 2, and Rl means a hydrogen atom or a hydroxy group, x2 meanS xl or a -(cH2)n-(O~l-(CH2)k (C6H4)q group wherein k means the numbers 0 to 4, 1 and q mean the number 0 or 1, and R2 means a hydrogen atom, a Cl-C4-alkoxy group, a functional group, . :
:..~. -.-~ : :

. . .- . ~
, 2034Z~42 or, boulld via this functional group, a bio- or macromolecule, A , B , B , C , C2, Dl, D2 El E2 F1 d 2 in each case X2, independently of one another, 5 G means R2 or a second macrocycle, bound via K, of gener~l Formula II

Q~ ' ZOOC-CH--N N--CH-COOZ III), ,~ 1 / \< D 2 D~ _. N ~ 2 D 1 CH _X2 COOZ ' wherein K means a direct bond, a bis(carbonyl- ~:
amino)group (-NH-CO-CO-NH-), or a Cl-C14-alkylene group which optional]y carries at ~: the ends carbonyl (>CO) or carbonylamino (-NH-CO-) groups or oxygen atoms, and which contains optionally one or several oxygen atom(s), hydroxymethylene (-CH-OH-), C~l(X2)COOZ-, acyl-substituted or hydroxy-acy}-substituted imino groups, or one to two C-C-double andtor C-C-triple bonds, Z is a hydrogen atom and/or a metal ion equivalent of an element of atomic numbers 21-29, 31, 32, 37-39, 42-44, 49 or 57-83, ~: ~

- , ~ , .. .
, :.
' Z0342~t2 witll ~he proviso that the 12 ring substituents throucJh F~ stand for at least 8 hydrogen atoms, that Xl and x2 stand simultaneously for hydrogen only iE at least one of the ring substituents A through F
does not mean a hydrogen atom, and that the macrocycle of genera]. Formula I contains no more than one bio-or macromolecule, and that, if desired, the remainder of the C02~l groups is present as an ester or amide, as well as their salts with inorganic and/or organic bases, amino acids or amino acid amides.

Preferred are tetraaza compounds of general Formula III

ZOOC-CH--N N CII-COOZ
/. \~ D 1 k ¦ ct C~l x COOZ

wherein . .
.
- - ~ ' .

Z0342l~2 --- is a single or double bond, ~ is a nitrogell atom or the residue NEI, X is a hydrogen atom, a -~C112)n-R group or a -(Cll2)n~- (fH) n-CE120EI group wherein n means the numbers l to 5, `
m means the numbers O to 2, and . -Rl means a hydrogen atom or a hydroxy group, x2 means Xl or a -(CH2)n-(0)l (CH2)k ( 6 4 q group wherein k means the numbers O to 4, ~ :
l and q mean the number O or l, and R2 means a hydrogen atom, a Cl-C4-alkoxy group, a functional group, or, bound via this functional group, a bio-or macromolecule, Al, Bl, Cl and D1, independently of one another, mean x2 in each case, G means R or a second macrocycle, bound via K, of general Formula IV

~ .
/!;;;.Q~\
1 1 1 1 l I
ZOOC-CII--N N CH-COOZ ~IV~

N ~ D I

Cooz .

X034;~2 w~ reill K means a dircct bond, a bis(carbonyl-a~ o)yroup (-NII-CO-CO-NII-), or a Cl-C14-alkylene group which optionally carries at the ends carbonyl (~ Co) or carbonylamino (-NII-CO-) groups or oxygen atoms, and which contaills optionally one or several oxygen atom(s), hydroxymethylene (-CI~--OH-), Cll(X2)COOZ-, acyl-substituted or hydroxy-acyl-substituted imino groups, or one to two C-C-double and/or C-C-triple bonds, Z is a hydrogen atom and/or a metal ion equivalent of an element of atomic numbers 21-29, 31, 32, 37-39, 42-44, 49 or 57-83, with the proviso that Xl and x2 simultaneously mean hydrogen atoms only if at least one of the 4 ring substituents Al, Bl, Cl and Dl does not stand for a hydrogen atom, and that, if desired, the remainder of the CO2H groups is present as an ester or amide, as well as their salts with inorganic and/or organic .
bases, amino acids or amino acid amides.

Compounds of general Formula I wherein Z means hydrogen are denoted as complexing compounds, and with ~ at least two of the substituents Z meaning a metal ion :~ equivalent, are called metal complexes.
The element of the above-mentioned atomic num-ber forming the central ion of the phy~iologically compatible compIex salt can, of course, also be radio-~ active for the intended purpose of using the diagnostic : agent of this invention.

.'. ~ , .
, .

_ 9 ~0;~2~t2 1ll C~IS~ the agent of the invenLion is intended for u~e in NMI~ diayllostics, the central ion of the complex s~lt must be paramagnetic. These are, in particular, the divalent and trivalent ions of the elements of atomic numbers 21-29, 42, 44 and 5~-70. Suita~le ions are, for exa~ple, the chromium(III), m~ng~nese(Il), iro~ ), cobalt(II), nickel(II), copper(II), praseodymium(III), neodymium(III), samarium(III) and ytterbium(III) ions. On account of their very strong magnetic moment, the gadolinium(III), terbium(III), dysprosium(III), holmium(III), erbium(III) and iron(III) ions are especially preferred.
For using the agents of this invention in nuclear medicine, the central ion must be radioactive.
Suitable are, for example, radioisotopes of the elements copper, cobalt, gallium, germanium, yttrium, strontium, technetium, indium, ytterbium, gadolinium, samarium and iridium.
If the agent according to this invention is intended for ~se in X-ray diagnostics, the central ion must be derived from an element of a higher atomic number in order to attain adequate absorption of the X-rays. It has been found that diagnostic media con-taining a physiologically compatible complex salt with central ions of elements of the atomic numbers between 21-29, 42, 44, 57-83 are suitable for this purpose;
these are) for example, the lanthanum(III) ion and the above-mentioned ions of the lanthanide series.
Preferred groups for Xl that can be mentioned are CH2OH, CH2CH2OH and CHOHCH2OH; and preferred groups for X2, Al, Bl, Cl and/or Dl are CH2OH, CH2CH2OH, CH20CH2C6H5, CHOHCH20H, CH2C6H40CH3 ~ CH2C6H5 ~
CH2C6H4O(CH2)3COOH, CH2C6H4NCS, wherein the remaining ring substituents A2, B2, c2 ~2 El E2 Fl 2 preferably mean hydrogen.

.

~03~ZLt 2 ~ alkylelle chaill stallding for K, to which the second m~lcrocycle II and IV, respectively, is linked, carries at the ends optionally carbonyl ~CO), carbonylamino (NII-CO) groups or oxygen atoms and contains 1-14 carbon atoms. This chain can be inter-rupted by one or several oxygen atom(s), hydroxymethyl-el1e (-CllOII-), CII tX2) COOr~rJ , acyl- or hydroxyacyl-substituted imino groups or one to two C-C-double and/or C-C-triple bonds. The two macrocycles can, however, also be linked by a direct bond. Suitable optionally hydroxylated acyl groups are acyl residues of up to 10 carbon atoms.
Examples in this connection are the acetyl, propionyl, butyryl, benzoyl and hydroxyacetyl residues.
The alkylene chain can be straight- or branched-chain, saturated or unsaturated, and can be interrupted, if desired, as described above. This chain can contain up to 4 oxygen atoms and/or up to 3 carboxymethylimino groups.

: .
.. .. . -: ' ' ~ ' ": ' ' .

20342L~2 Exampl~s ~or the alkylene chain are: .

2 2 2 2 ' 2 ) ~ . ~ CH2 C~t2- o- CH2- Cl~z 1 ~ - ~ CH2- O- CH2 ) 2 2 3 ' 2 Cl~z lo C112-CH2~3-, -C~l2-cH2-lo-cH -CH ) -O O
Il 11 -CH-CH-, -CH2-CIl-lCH-CH2-, -C-C-C_C-, -NH-C-lCHz)o 6-C-NH

O O
2 1 2 C1~2 1 ~H2 C~2 1 CH2-- -C-lcl12)l 6-C-, -O-(CH2)~ 6--~
CHX CHX CHX
Icooz Icooz Icooz -C~-N-C~12- , -1CI12~2CH C~1 1CH212 :
.

, , :' ' '' . ,' ' . . ' , . , ~ ' ' ' '' ' ' '`.
, '' ' ~' . ' ,' '' ' , , . ~' ., ' , .' ' ' " , ~ . , ~ . , , '" ' ' ~ , ' ' I
,, ~

- 12 - ~0342L12 Pre~erably, the two m~ro~yales are l~n~ed by dire~t bond or by the group -(O)q~(CH2)n~( fH) ~-OH
~ 2),~ ()1-~he functional groups ~re any molety capable o~
rea¢~lng in a ~anner wh1~h will permi~ attachment o~ a biomole~ule or macromolecule.
Pre~er~ed ~unctional ~roups which R2 can repres~nt are, for example, the maleimidob~nzoyl, 3-sulfomAleimidobenzciyl, 4-(1nalc:imidomethyl)~y~3~hexyl-carbonyl, 4 - t 3 -~ul f o- (mal eimidome~hyl ) ~ cyal ohoxylaarbonyl, 4- (p-maleimitlopll~nyl) butyryl, 3- ~2-py~idyldithlo1 proplon-yl, me~hacryloyl~pent~methylen~amido, bromQa~etyl, ~ -iodoacetyl, 3-iodopropyl, 2-bro~o~tllyl, 3-mercaptopropyl, 2-mercaptoethyl, phen~lenelsothio~yanate, 3-aminop~opyl, benzyl es~er, ethyl ester, te~t bu~yl este~, a~ino, hyd~oxy, Cl-C6-alkylaminc~, aminoc~bonyl, hydrazino, hydrazinocarbonyl, maleimido, methac4ylamido, methacryloylhydrazinocarbonyl, maleimidamidocarbon~l, halogeno, mercapto, hydra~inot~imethylen~hydrazino-çarbonyl, aminodimethyleneami~oearbonyl, bromo~rbonyl, phenylenediazonium, isothiocyanRte, semic~rb~zide, thiosemicarbazide, isocyanate groups.
Several s~lected groups will ~e sct forth 2S for the sake of ~xplanat1On:
n o -C~12-C6H~OlCH2I~-H~[I , -CH2~C61~-OlCHzl~NH~ .NN-N~
o cn2-c6~ olch2~5cozcH?c6H5~ ~l 2 C6~ -CNt-c2cH2c6Hs~
o -CH2-C6H~ o~CH2~scorJHNN2i CHt CCH~ CH2-C6~ 01CH2~-5N, Chz~c6H~olchzl~NNNH2~ C 2 6 ~ 2 $

-C:H2-C~,H~-O ~C:Hz) ~Br, -COz-C6H~-NO2~ ' , . ~

, .
.: , :
~ ~ .
' - 1 3 - 20342~Z

Cll C tl 01 CH ) CONHNII- I C112 ) ~-N\INH2 ~ -C112-NIIN112 ~ 2 2 2 2 3 C~12 CGH~ O CH2C8r -c6~N~lcoctl2Eir 2 6 ~ 2 2)2N~2 C~2-C61i~-N1~2- -C6~,.-N2~ -C6H~NC5 -CH2-C6tl~-N11-C-lc~12~2-S-s- ~1 -Nl~co-Nl~-NH2~ -N~CS NH NH2 -C~2-C6H~-O-CH2-cl~-ctl2~ -Ct~2-c6Hl-o-cH2-c-NH-lcH2llo-c-NHNH2~

-CH C H -O-CH -C11011-C\1 -NHICH ~ o-C-NHNH2~ -OCH2-C-N-CH2-lCHOH),-CH20H

2 2 2 2 3 ~ ¦ 2 2 -. .

-CH2-0-1CH21.~-NHNH2, -CH2-0-CH2-C-NH-NH2, -CH2-0-CH2-CH2-NH2, -c~2-o-c-2-NH-c- ~H2~2-S-S ~ ' CH2--CH2-C-NH- ~CH2~ lo-C-NH-NH2, C H - N/ ¦~ - C-C - C_C - R - C_C - C H: C R R

O
O
O O~11_ -CCH2Eir -C-c6H~o C 2 ~.~¦¦ -NH-Co-cH2-l~r~ -NH-CO-CH2Cl O

;~

: ., . , . ` . ~ -' ~ ' ' : ~ ' '`' ~034Z' ¦ Z

C 11 Cll -N ~ C11!13-C6H~-N~ C CH~ ~ 2 3 O ~N~
2)~N~12, -C~,H,,SCN, -CCICH3~=CH2, -C~CRR , C6H~,CH2Br, 05 i ( C ~3 I
I CRR' -CH Br -CH2~, -C~=CH-CH2Br, -OS02C6 ~, ~ 2 -IC-C1 -C-O-C-R -C-OR, -C-N, -C-N ¦, -cH=cH-co2R~

wherein R and R' are identical or different and in each case mean a hydrogen atom, a saturated or unsaturated Cl-C20-alkyl residue optionally substituted by a phenyl group, or a phenyl group. The -NCS, -NO2, -OH, -NHNH2,-NHCOCH2Br, -NHCOCH2Cl, -CO2H and -CON3 groups are especially preferred.
The residual acidic hydrogen atoms, i.e. those that have not been substituted by the central ion, can optionally be replaced entirely or in part by cations of inorganic and/or organic bases or amino acids.
The corresponding acid groups can also be entirely or partially converted into esters or amides.

- .
.

X03~2~12 ~ <~ inorgallic cations ar~, for example, t:ilC 1 i ti~i.UIll iotl, ~he potassium ion, tlle calcium ion, !:he mclc3n~xiull~ iOI~ and especially the so~ium ion.
',ui~abk cations of organic bases are, inter alia, those of l)rimary, s~condary or tertiary amines, such as, for ex~mple, ~h~lnolamille, diethanolamine, morpholine, glucamin~, N,N-dimethylglucamine, and, in particular, N-methylglucamine. Suitable cations of amino acids are, for example, those of lysine, of arginine, and of ornithine, as well as the amides of otherwise acidic or neutral amino acids.
Suitable esters are preferably those with a Cl-C6-alkyl r~sidue; examples that can be mentioned are the me-thyl, ethyl and tert-butyl, benzyl and 4-methoxy-benzyl residues.
If the carboxylic acid groups are to be presentat least in part as amides, then suitable residues are saturated, unsaturated, ~traight- or branched-chain or cyclic hydrocarbons of up to 5 carbom atoms which are optionally substituted by 1-3 hydroxy or C1-C4-alkoxy groups. Examples that can be mentioned are the methyl, ethyl, 2-hydroxyethyl, 2-hydroxy-1-(hydroxymethyl)ethyl, l-(hydroxymethyl)ethyl, propyl, isopropenyl, 2-hydroxy-propyl, 3-hydroxypropyl, 2,3-dihydroxypropyl, butyl, isobutyl, isobutenyl, 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl, 2-, 3- and 4-hydroxy-2-methylbutyl, 2-and 3-hydroxyisobutyl, 2,3,4-trihydroxybutyl, 1,2,4-trihydroxybutyl, pentyl, cyclopentyl and 2-methoxyethyl groups. The amide residue can also be a heteracyclic 5- or 6-membered ring formed with the inclusion of the amide nitrogen. Examples that can be cited are: the pyrrolidinyl, piperidyl, pyrazolidinyl, pyrrolinyl, pyrazolinyl, piperazinyl, morpholinyl, imidazolidinyl, oxazolidinyl, thiazolidinyl rings.

Z0342~t2 Th~ comp~und~ of this inven~ion e~hibit ~he deslred prop~rties described in thc for~oing. ~hs ¦
complexe~ contain ~he meta~ ion~, requi~ed ~or thair u~age, ~tably bound t~erein.
The valu~ of the o~molality, responsible for side efEects, sucll a& pa~n, damage ~o the blood vessels, and cardio~ascular disturbances, ls markedly reducea as compared with MAGNEVIS~ (compare Example lb: 0.55 ~osmol/kg] with ~A~VIST~ 1.96 {o~mol/kg~, O.5 mol/1 at ;
37 ~) Th~ valuc ~or the magnitude of rel~xivity, ~epresen~ing a m~u~e for imaging in MRI, is sur-pris~nyly high ~he signal amplifica~ion in the pla~ma could 4e increased, for example in the ca~e o~ the compound lS of Exampl~ lb, by ~wice the value afi compared witl MAG~EVIST~.
A furthcr ~dvantage of tho present inven~ion residcs in that complexes with hyd~opl-ilic or lipophilic substituents have now become accessible. Thi~ affords th~ possibility of con~rollin~ the compatibility and pharmacokine~ics of ~hese complexes by chemical substitutiol~ ~
sy ~he choice of suitable bio- or macro~olecules lsee arther below) in R2, co~plexes of this inven~ion are obtain~d ~hich exhlbi~ a surpri~in~l~ high tlssue and or~an specificity.

,,, , : .

. ! , :

- 1~ 2034X~2 ; The compounds o~ thi~ inv~ntion are y~odu~d by ~plittlng~f the blocXin~ group~ conven~onally i~
compounds of ~neral Formula I' a /'`; 4 ZO -CN ' 1~ N CH, I:OOZ

N ~ D

Cll _X2 !o~

wherein G', Xl and x2 in each case stand for G, Xl and X2, the hydroxy group~ and functional groups ~ontain~d therein being present in the blocked form and, regpectively, A9 a precursor, and Z' means a hydrogen atom or an acid blockinggroups : optionally g~ne~ating the desi~ed functional group;
reacting, if desired, the thus-obt~ined co~plexing compounds o~ general Formula I wherein Z me~ns hydrogen in a mannor known per s~ wlth at le~st one metal oxi~c o~ ~ctal salt of an el~men~ o~'a~omic nun~bers 21-29, 31, 32, 37-39, 42~44, 4~ or 57-83 ~ optionally binding the :function~1 ~roups to a bio-: ~ or macromoleculc -- whereln the complexing ~an take place before o~ after thc split~lnq of o th~ blocklng g~oups ~or the ~ydroxy gr~ups and functional qroup~
:
.
i : ,.

.03~2'~2 a~d/~r generation ok the ~unatior-al groups and link~ge ~o ~ m~ro- ~ biomoleculQ -- and subsequen~ly, if d~3sire~ bstituting any still present ~cidi~ hydrogen atoms by cations of inor~anic and/or organic b~e~, amino acids or amlno acid amides or, respec~ively, converting the corresponding acid groups entirely or p~rtially into est~rs or amides.

Suitable acid ~locking g~oups Z' are lower alXyl, aryl and aralkyl ~roups, e.q., the me~hyl, ethyl, propyl, n-hutyl, tert,bu~yl, phenyl, ben~ylr d$phenyl-me~h~l, triphenylmeth~l, bis(p-nltrophenyl~methyl groups a~ well as trialkyl~ilyl groups.
Z' can also stand for an alkali metal.
The bloekin~ groups are spli~ o~f iJl accord-allce with methods known to tho~e skille-l in the art, for ex~mple by hydrolysis, hydrogenolysis, ~lk~line saponifica~ion of tlle este~s with alkali in an aqueous-alcoholic ~olution at t~3mperature~ of ~ to 50 C, acidie ~Aponif ication wi~h mineral acid~ o~, in ~a~e 20 of ~ert. butyl ester~, for example, with ~he aid of tr i f 1 uoroacet ic ac id .
Suitable hydroxy blocking groups are, for ~xample, the ben~yl, 4-1nethoxyben~yl, 4-nit~oben~
trityl, dipheny~methyl, trimethylsilyl, dimethYl-tert.- :
2S butylsilyl, diphenyl-tert.-butyl~ilyl gro~ps.
The hydroxy group~ can also be pr~3CI~t~ f.or example, a~ tetrahydrophyranyl (THP) ethers, ~-alkoxy~thyl ethers, 2-methoxy-e~hoxym~hyl ~MEM) ethers, or as ester~ with a~omatic or aliphatic carboxylic acids, such a~, for example, acetic aai~ or benzoic aaid. ~n case of polyol~, th~ hydroxy group~ aan also be block~d ~n ~he form of Xe~alg with, for example, ac~tone, ace~aldehyde, cy~lohexanone or benzaldeh~de.

Z0342~2 1' 2' The llydroxy groups present in X and X
can also be present in blocked form by intramolecular esterification with the ~-positioned carboxy groups ~o obtain the corresponding lactones.
The hydroxy blocking groups can be liberated in accordance with literature methods known to a person skilled in the art, for example by hydrogenolysis, reductive cleavage with lithium/ammonia, acid treatment of the ethers and ketals, or alkali treatment of the esters (see, for example, "Protective Groups in Organic Synthesis", T.W. Greene, John Wiley and Sons 1981).
The synthesis of dimeric compounds, i.e.
compounds containing a second macrocycle of general Formula II or IV, takes place according to methods dis-closed in the literature, for example by way of anaddition/elimination reaction of an amine with a carbonyl compound (e.g. acid chloride, mixed anhydride, activated ester, aldehyde); of two amine-substituted rings with a dicarbonyl compound (e.g. oxalyl chloride, glutaric dialdehyde); of two p-nitro-substituted nitroxides with bisalcoholates [cf. E. Klingsberg, The Chemistry of Heterocyclic Compounds, Interscience Publishers New York, -p. 154 (1961)1, of two rings each exhibiting a nucleo-philic group, with an alkylene compound carrying two leaving groups or, in case of terminal acetylenes, by oxidative coupling (Cadiot, Chodkiewicz in Viehe "Acetylenes", 597-647, Marcel Dekker, New York, 1969).
The ring-linking chain can subsequently be modified by secondary reactions (e.g. hydrogenation).
The synthesis of directly linked compounds (i.e. K meaning a direct bond, see Example 4) can be accomplished by cyclization of tetrahalogenomethyl-4,4'-bispyridines (see fuFther below).
~ '' .

, :
:: :

~034ZL~2 Tllc <;ynt:tlesis of the educts I' takes place !~y alkylation of compounds of gencral Formula V
F 1 /~.~ j~l! I

u ---N N--u (V), D 2 ¦ C

wherein 5 U is hydrogen and V is an amino blocking group or U represents amino blocking groups and V is hydrogen, wherein U and V can also be identical, with compounds.of general Formula VI
x1 ~' Nf-CH-COOZ IV~ ) or with compounds of general Formula VII
x2 Nf-CH-COOZ ~VI 11 ~

wherein Nf stands for a nucleofugal entity, such as, 15 for example, Cl, Br, I, CH3-C6H4S03, CH3S03, 4-N02-C6H4s03~ CF3S03-" .

. .. .

, , :
, - 2~ - ~03~2~2 ~ r}~e hydroxy groups that may be c~ntained in xl and x2 can A130 form a l~tone together wLth the OZ' resi~ue Ex~ples for alkylating re~g~ntC ~re~ bro~o-aceti.c acid, c~lloroacetic acid, bromo~cetic acidme~hyl ester, ~romo~ce~ic acid ter~,butyl estex, chloroacetic acid benzyl ester, 2-cllloro-3~benzyloxy-propanoic acid sodium ~alt ~P 0,32$,762), 2-bromo-3-b~nzyloxypropanoi~ acid tert-butyl est~r (J. Gen. Chem., USSR 36 : 52, 1966), 3,4-o-isopropylidene-~-p-~olyl-sulfonyl-3, 4-dihydroxybutyric acid ethyl este~ ~Synth.
Comm. lg : 3077, 19~9), a-bromo~y -butyrolaetone.
Examples of amino ~locking groups U and ~, resp~ctivel~, are: formyl, ~rifluoroacetate, ~enzoate, 4-ni~robenzoate, a~e~ate, tosylate, mesylate, benzyl, 4-nit~obenzyl, 4-mettloxybenzyl, tritnethylsilyl, dimethyl-tert,butylsilyl.
Alkylation of the compounds of ~eneral For-mula v to the educts o~ general ~ormula I' with the ~ompounds of general Formula VI or VII ~akes place in pola~ aprotie ~olvents, su~h as, for example, dime~hylformamide, acetonit~ile, dimethyl sulfoxide, ~queous tetrahydrofuran, dioxane, or hexamethylphosphoric tria~ide in the presence of an a~id c~ptor, suoh as, :~5 cor example, ~erti~ry amlne (e.g., triethylamine, trimethylamine, N,N-dimethylaminopyridine), 1,5-diazabicyclol4.3.0JnQnene-5 ~D~N), 1,5-diaza~i-cyclo~5.4.03undecene-5 tDBU), alkali, alkal~ne earth car~onate, bicar~onate or hydroxide ~g., sodium, lit~ium, magnesium~ calcium, bariu~, pota~lum oarbonate, hydroxide and bicarbonate) at temperatures of between -lO C and 120-C, preferably be~ween O^C and 50-C, it being possible ~o ad~, if de~ired, cataly~ic ~moUnts of iodide or bromide.

- ~
-.

203~2L~2 ~ ft~ splitting off the remaining amino block-ing group(s) according to methods known to one skilled in the art ~for example acidic or alkaline hydrolysis, hydrogenolysis, reductive cleavage with alkali metals in liquid ammonia, reaction with tetrabutylammonium fluoride), the remaining amino function(s) is or are reacted in a second alkylating reaction with VII and VI, respectively, so that compounds are obtained wherein Xl ~ X2.
The conversion of a precursor of the desired 6-membered ring contained in the final product takes place according to methods known to persons skilled in the art. Examples worth mentioning are the hydrogenation of pyridine ~Advan. Catal. 14 : 203 (1963)], deoxygena-tion of nitroxide rings [E. Klingsberg, The Chemistry of Heterocyclic Compounds, vol. 14, part 2, Interscience Publishers New York, p. 120 (1961)1, conversions and introduction of functional groups at the 6-membered ring, e.g. liberation of phenolic hydroxy groups ?0 lJ. Org. Chem. 53 : 5 (1988)], introduction of halogen substituents [E. Klingsberg, The Chemistry of Hetero-cyclic Compounds, vol. 14, part 2, Interscience Publishers New York, p. 341 (1961); Houben-Weyl, Methoden der organischen Chemie, vo. V/3 : 651 (1962)1.
Functionalization of 4-halopyridine derivatives (e.g. azide exchange) in the phase transfer process with the use of 18-crown-6 or tetrabutylammonium halogenide as the catalyst has been disclosed in "Phase Transfer Reactions" (Fluka Compendium vol. 2, Walter E. Reller, Georg Thieme publishers, Stuttgart, New York). A thus-obtained azide group can be con-verted into an amino function by using methods known to one s~illed in the art (for example, catalytic .
: , . . - . .

, - . . . -:
:, ;. ' ,....... . ' - :

- 23 - ~03~2~2 ~Iy~lr~3~nation~ uben-Weyl, "Methoden d~r organischen Chemi~" v~l. 11/1 : 539) or reaction with Raney nickel/
hydr~zine (German Patent Application 3,150,917~. This amino function can be transformed into an isothio-cyan~te group by means of methods known from theliterature (e.g. with thiophosgene in a two-phase system, S. Scharma, Synthesis 1978 : ~03; D.K. Johnson, J. Med. Chem. 1989, vol. 32, 236).
~y reacting an amino function with a haloacetic acid halogenide, an -haloacetamide group can be generated (JACS 1969, vol. 90, 4508; Chem. Pharm. Bull.
29(1), 12~, 1981), which is suitable, just as the iso-thiocyanate group, for example, for coupling to bio-and macromolecules.
The synthesis of the compounds of general Formula V is conducted by cyclization according to methods known from the literature [for example, Org.
Synth. 58 : 86 (1978), Macrocyclic Polyether Syntheses, Springer Publishers Berlin, Heidelberg, New York (1982);
Coord. Chem. Rev. 3 : 3 (1968); Ann. Chem. 1976 : 916;
J. Org. Chem. 49 : 110 (1984)]; one of the two reactants carries two leaving groups at the chain end, the other carries two nitrogen atoms which displace these leaving groups in nucleophilic fashion.
An example that can be cited is the reaction of Al-D2-substituted diethylenetriamines, the terminal-positioned nitrogen atoms of which displace, in nucleo-philic fashion, the leaving groups of, for example, 2,6-dihalomethylpyridines, 2,6-ditosylmethylpyridines or 2,6-dimesylmethylpyridines. For the synthesis of directly linked dimers, 2,2',6,6'-tetrachloromethyl-4,4'-bispyridines (see, for example, Synthesis 552, 1989) are utilized in the cyclization reaction.

~,: , . -:-X03~ Z
- 2~ -The nitrogen atom~ are optionally blocked (fo~
ex~mple as t~ylates or trlfluoroacetates) and are liberated ~rior to the subsequen~ alkylatin~ reac-tion in accordance wi~h me~hods kno~n ~rom the litera-tyre tthe tosylates, for example, with min~ral acids,alkali ~e~als il~ llqui~ ammonia, hydxQ~rom~c acid and phenol, REDAL~ (80dium-bl~ methoxyethoxo)-aluminum-dihydxide~ thium aluminu~ hydrlde, ~odiu~ amalqam, compare, for example, Liebigs Ann. Che~. ~977, 1344:
Tetrahedron Let~er3 1~: 3477: the trifl~oroaaetat~, for ex~mple, w~th min~ral acld~ or ammonia in ~othanol, compare, for example, Tetrahedron Lettcr~ 1967: 289).
In order to pr~pare macrocycles w~h differ-ing substitution on the nitrogen atoms, these a~oms can be provid~d, in the educts, with dif~erent blocking groups, for example with tosylate and bcnzyl groups.
The la~ter are then likewise removed according to con-ventiot)al methods disclosed in the l~teratu~re (preferably by hydrogcnation, e.g.~EP Patent Applica-tion ~32,7~1).
In case diesters are utilized in the cycliza-tion rcaction, the resultan~ diketo compounds must be reduced according ~o methods known to one skilled in the art, for example with dibor~ne.
It is also possible to cyclize correspondingly substituted termin311y positioned bisaldehydes or bi~-ketones, e.g.~2,6-bisacetylpyri~ines, with the re-spectively de~ired tor~inal-positioned bisami~e~ the reduc~ion of ~he ~hus obt~ined Schiff bases takes 3~ place accord~ng to methods known ~rom the ll~e~atu~e, e.gJ by catalytic h~drogena~lon ~11elv. Chim. ActA 61 : -~
1376 (1978~l.

. .
: . , , - .
. . . .
.

.
~ ' , , ` .
, . , '-: ~

.

- 25 - ~0342 ~ amil~es requiLed as star~in~ materials for th~ cyclization are produced in analogy to methods known from the literature ~e.g., EP 299,795). Starting with an N-blocked amino acid, a triamine is obtained by reaction with a partially blocked diamine (for example accordiny to the carbodiimide method), splitting off of tll~! blockillg CJroUp5, and diborane reduc~ion.
Suitable substituents convertible into the functiollal group that can be linked to a macro- or bio-molecule are, inter alia, hydroxy and nitrobenzyl,hydroxy and carboxyalkyl, as well as thioalkyl residues of up to 20 carbon atoms. They are converted, accord-in~ to literature methods known to one skilled in the art ~Chem. Pharm. Bull. 33 : 674 (1985); Compendium of Org. Synthesis vol. l-5, Wiley and Sons, Inc.; Houben-Weyl, Methoden der organischen Chemie, vol. VIII, Georg Thieme publishers, Stuttgart; 3. Biochem. 92 :
1413 (1982)], into the desired substituents (for ex-ample with the amino, hydrazino, hydrazinocarbonyl, epoxy, anhydride, methacryloylhydrazinocarbonyl, maleimidamidocarbonyl, halogeno, halogenocarbonyl, mercapto, isothiocyanate group as the functional group). In case of the nitrobenzyl residue, catalytic hydrogenation (e.g. according to P.N. Rylander, Catalytic Hydrogenation over Platinum Metals, Academic Press 1967) to the aminobenzyl derivative must first be performed.
Examples of conversion of hydroxy.or amino groups linked to aromatic or aliphatic residues are the reactions performed in suitable solvents, suçh as tetrahydrofuran, dimethoxyethane or dimethyl sulfoxide, two-phase aqueous systems, such as, for example, water/dichloromethane, in the presence of an acid captor, such as, for example, sodium hydroxide, sodium hydride, Z0342~1Z

or a~k~li or alkaline ea~t~ ~arbonate~, such as, ~or example, .sodl~m, ma~ne3ium, pota~ lum, calcium carbona~e or poly(4-vinylpyridine) REILLEX~, ~t temperature~ between 0 C and the boiling poin~
of the re~pective solvent, preferably, however, between 20 C and ~0" C, with a ~ubstrate o gener~l Fo~mula VIrI

Nf - L - Fll (VIII~ :

wherein ~: , L means an aliphatic, arom~tlc, arylaliphatic, bxanched, straigh~-chaln or cyclic hydrocarbon residu~ of up to ~0 carbon atoms, and Fu means the desired terminal-positioned functional g~oup, optionally in the ~lo~ked form (DOS ~,417,413~
Examples of compounds of general ~ormula VIII
that can be cited are:

3rlCH2)2~1Hz, ~r~cH2~3oH~ BrCH!COOCH3, BrCH2C~zt-Bu, ClCHzWNHNHz~

OrlCH~ COzC2H~ OrCH2CO~r, OrCH2CO~H2, ClC~12COOC2H5, ~ \
~rCHzC~NHNH2~ OrC1~2-CH-CH2. CF~SOlICHz1~9r, OrCN2C CH, OrCN~Ch~CH2, OrcH2C6H~cs -, ' ' ~. ', ' .'' :
... . . .. .

~0342L~ 2 Conversions of carboxy groups can be con-ducted, for example, according to the carbodiimide method (Fieser, ~eagents for Organic Syntheses 10 : 142), by way of a mixed anhydride [Org. Prep. Proc. Int. 7 :
215 (1975)], or via an activated ester (Adv. Org. Chem.
p~rt B, 472).
The thus-obtained complexing ligands (as well as the complexes~ can also be linked to bio- or macro-molecules of which it is known that they are partic-ularly accumulated in the organ or organ part to be examined. Such molecules are, for example, enzymes, hormones, polysaccharides, such as dextrans or amyloses, porphyrins, bleomycins, insulin, prosta-glandins, steroid hormones, amino sugars, amino acids, peptides, such as polylysine, proteins (e.g. immuno-globulins, monoclonal antibodies, lectins), lipids (also in the form of liposomes), and nucleotides of the DNA or RNA type. Especially to be emphasized are conjugates with albumins, such as human serum albumin, antibodies, e.g. monoclonal antibodies specific for tumor-associated antigens, or antimyosin. In place of biological macromolecules, it is also possible to use suitable synthetic polymers for the linkage, such as polyethylenimines, polyamides, polyureas, poly-ethers, such as polyethylene glycols and polythioureas.The pharmaceutical agents produced therefrom are suitable, for example, for use in tumor and inarction diagnostics as well as tumor therapy. Mono~clonal antibodies (e.g. Nature 2S6 : 495, 1975) show the advantagesover polyclonal antibodies that they are specific for an antigenic determinant, possess defined binding affinity, are homogeneous (thus substantially simplifying their preparation in pure form), and :-. . :
: . . :
: ~ . .
: :
: . ' ' ,: .' - ~3 -- ~
~034X~2 call ~e ~roduce~ ell cul~ures in large amounts.
Suitab]e as sucl~ are, ~or example for tumor imaging, ~onoclonal antibodies and/or their fraqments Fab and F(ab')2 specific, for example, for human tumors of the ~astroin~estinal tr~ct, the breast, the liver, the bladder, the gonads, and melanomas ~Cancer Treatment l~epts. 68 : 317 tl984); siO. Sci. 34 : 150 (1984)], or act- -ingagainst carcinoembryonic antigen (CEA), human chorionic gonadotropin (~-EICG), or other tumor-positioned antigens, such as glycoproteins [New Engl. J.
Med. 298 : 1384 (1973), U.S. Patent 4,331,647].
Suitable are also, inter alia, antimyosin, anti-insulin, and antifibrin antibodies (U.S. Patent 4,036,945).
Colon carcinomas can be detected with the aid - -of conjugates, complexed with gadolinium(III) ions, with the antibody 17-lA (Centocor, USA) by means of NMR diagnosis.
Suitable for liver examinations and, respect-ively, for tumor diagnosis are, for example, conjugates or inclusion compounds with liposomes which are used, for example, as unilamellar or multilamellar phosphatidylcholine-cholesterol vesicles.
In case of the antibody conjugates, binding of ~:
the antibody to the complex or ligand must not lead to loss or reduction of binding affinity and binding specificity of the antibody to the antigen. This can be accomplished either by binding to the carbohydrate portion in the Fc part of the glycoprotein and/or in the Fab or F(ab')2 fragments, or by binding to sulfur atoms of the antibody or, respectively, the antibody fragments.

- ~.. - , , , .

- 29 - X03~Z~

lil tll~ rirst inst~nce, an oxidative cleavage o~ su-~al^ ullit:s mus~ ~irst be per~ormed for the genera-~ion of ormyl groups cap~ble of coupling. This oxid~ltion can be carried out by chemical methods with oxidiæinc3 a~ents SUCll as, for example, periodic acid, sodium mc~tcll~eriodate, or potassium metaperiodate in accordance with methods known from the literature (e.g., J. I~istochem. and Cytochem. 22 : 1084, 1974) in an aqueous solution in concentrations of l - lO0 mg/ml, preferably l - 20 mgtml, and with a concentration of the oxidizing agent of between 0.001 and lO millimoles, preferably 1 to lO millimoles, in a pH range of abou-t 4 to ~ at a temperature of between 0 and 37 C
and with a reaction period of between 15 minutes and 24 hours. The oxidation can also be performed by enzymatic methods, for example with the aid of galactose oxidase in an enzyme concentration of 10 - 100 units/ml, a substrate concentration of 1 - 20 mg/ml, at a pH of 5 to 8, a reaction period of l - 8 hours, and a temperature of between 20 and 40 C (for example, J. Biol. Chem. 234 : 445, 1959).
Complexes or ligands with suitable functional groups, such as, for example hydrazine, hydrazide, hydroxylamine, phenylhydrazine, semicarbazide and thiosemicarbazide, are bound to the aldéhydes generated by oxidation; this is done by reacting between 0 and 37 C with a reaction period of l - 65 hours, a pH of between about 5.5 and 8, an antibody concentration of 0.5 - 20 mg/ml, and a molar ratio of the complexing compound to the antibody aldehyde of 1 : l to lO00 : l.
The subsequent stabilization of the conjugate takes place by reduction of the double bond, for example with sodium borohydride or sodium cyanoborohydride; the reducing agent is utilized herein with a 10- to lO0-35 fold excess (e.g. J. Biol. Chem. 254 : 4359, 1979).

. .

' .
' ' '':
: , .
.
.

~ 3() ~ ~03~2 ~2 Tlle second possibility of forming antibody COll jU~ateS starts with a gentle reduction of the disuLfide bridges of the immunoglobulin molecule;
itl this process, the more sensitive disulfide bridges between the l~ chains of the antibody molecule are cle~aved whereas the S-S bonds of the antigen-binding region rcmain intact so that there is practically no reduction in binding affinity and specificity of the antibody (~iochem. 18 : 2226, 1979; Handbook of Experimental Immunology, vol. 1, 2nd ed., Blackwell Scientific Publications, London 1973, chapter 10).
These free sulfhydryl groups of the inter-H-chain regions are then reacted with suitable functional groups of complexing compounds or metal complexes at 0-37 C, a pH of about ~ - 7, and a reaction period of 3 - 72 hours with the formation of a covalent bond which does not affect the antigen binding region of the antibody. Suitable reactive groups are, for example: haloalkyl, haloacetyl, p-mercuribenzoate, isothiocyanate, thiol, epoxy groups, as well as groups to be subjected to a Michael addition reaction, such as, for example, maleinimides, methacrylo groups (e.g. J. Amer. Chem. Soc. 101 :
3097, 1979).

.

~ . . , :
, . . . . ~ ~
: .. . ..

. ~

- 3~
~0342~Z
~ d~itionally, for linking the antibody fragments Witll the polymer complexes or with the ].igands, there is a number of suitable bifunctional "linkers" which are frequently also obtainable commercially (see, for example, Pierce, Handbook and General Catalogue 1986) which are reactive with respect to the Sll groups of thc fragments as well as with respect to the amino or hydrazino groups of the complexes.

Examples that can be cited are:

m-maleimidobenzoyl-N-hydroxysuccinimide ester ~MBS), m-maleimidobenzoyl-N-sulfosuccinimide ester (Sulfo-MBS), 5 N-succinimidyl-~4-(iodoacetyl)amino]benzoic acid ester ~SIAB), succinimidyl-4-(N-maleimidomethyl)cyclohexane-l-carboxylic acid ester (SMCC), - ~

- 32 - '~ 2~Z

succinimidyl-4-tp-maleimidophenyl)butyric acid ester (SMPB), N-succinimidyl-3-(2-pyridyldithio)propionic acid ester ~SDPD), 4-[3-(2,5-dioxo-3-pyrrolinyl~propionyloxyl-3-oxo-2,5-diphenyl-2,3-dihydrothiophene-1,1-dioxide, `
acetylalanylleucylalanylaminobenzyl, acetamido-p-thioureidobenzyl.

It is also possible to utilize bonds not of the covalent type for coupling purposes wherein ionic as well as van der Waals and hydrogen bridge bonds can contribute toward the linkage in varying proportions and strengths (key and lock principle) ~for example, avidin-biotin, antibody-antigen~. Also inclusion compounds (host-guest) of relatively small complexes in relatively large cavities in the macromolecule are possible.
The coupling principle resides in first producing a bifunctional macromolecule by either fusing an antibody hybridoma directed against a tumor antigen with a second antibody hybridoma directed against a complex according to this invention, or linking the two antibodies chemically via a linker ~e.g. in the way set forth in J. Amer. Chem. Soc.
25 lOl : 3097, 1979) or binding the antibody directed against the tumor antigen to avidin (or biotin, re-spectively), optionally via a linker [D.J. Hnatowich et al., J. Nùcl. Med. 28 : l294 ~1987)]. In place of the antibodies, it is also possible to employ their corresponding F(ab) or F(ab')2 fragments.
For pharmaceutical usage, first the bifunctional macromolecule is injected wbich is accumulated at the target site, and then, at a time interval, the complex .~ :
, - , , - : : - . : ~
- . : . :~ ,: . - : ~ -~ ~ ' . ' . ''.
,: ,. : -:: . . - ' ~ :

~0;~42~1i2 compound of this invention is injected [optionally bound - to biotin (or avidin)] which is coupled on at the target site in vivo and there can deploy its diagnostic or therapeutic activity. Moreover, other coupling ~ethods can likewise be utilized, such as, for example, "reversible radiolabeling"
described in Protein Tailoring Food Med. Uses [Am. Chem. Soc. Symp. 3~9 tl985)1.
A particularly simple method for the produc-tion of antibody conjugates or antibody fragment con-jugates is available in the form of the so-calIed solid phase coupling procedure: The antibody is coupled to a stationary phase (e.g. an ion exchanger) located, for example, in a glass column. By successive flushing lS of the column with a solution suitable for generation of aldehyde groups, washing, rinsing with a solution of the functionalized complex, and finally elution of the conjugate, very high yields of conjugate -are obtained.

This procedure permits the automatic and continuous production of any desired quantities of conjugates.
Also other coupling steps can be performed in this way.
Thus, for example, fragment conjugates can be prepared by the sequence of papain reduction/bi-functional linker/functionalized complex or ligand.
The thus-formed compounds are subsequently purified preferably by chromatography by way of ion exchanyers on a fast protein liquid chromatography unit.
,.

~., .

- , , - . .
~ , .

~034ZL~2 rrhc r~ al complexes of this invention are produc~d as disclosed in German Laid-Open Application 3,401,052 by dissolving or suspending the metal oxide or a metallic salt (e.g. the nitrate, acetate, carbonate, chloride or sulfate) of the element of a~omic numbers 21-29, 42, 44, 57-83 in water and/or Ln a lower alcohol (such as methanol, ethanol or isopropanol), and reacting with a solution or sus-pension of the equivalent amount of the complexing ligand and subsequently, if desired, substituting any acidic hydrogen atoms present by cations of inorganic and/or organic bases or amino acids.
Introduction of the desired metal ions can take place in this process before as well as after the splitting off of the blocking groups for the hydroxy groups and functional groups, or, respectively, before or after the generation of the functional groups and linkage to a macro- or bio-molecule.
Neutralization of any free carboxy groups still present takes place with the aid of inorganic bases (e.g. hydroxides, carbonates, or bicarbonates) of, for example, sodium, potassium, lithium, magnesium or calcium and/or organic bases, such as, inter alia, primary, secondary and tertiary amines, e.g. ethanolamine, morpholine, glucamine, N-methyl- and N,N-dimethylglucamine, as well as basic amino acids, such as, for example, lysine, arginine and ornithine, or of amides from originally neutral or acidic amino acids.
In order to prepare the neutral complex compounds, it is possible, for example, to add to the acidic complex ~alts in an aqueous solution or suspension such an amount of the desired bases that the neutral point is obtained. The resultant solution ,, ~

.
' ~

^~0;~2'12 ca~ then be concentrated to dryness under vacuum. It is frequently advantageous to precipitate the thus-formed neutral salts by adding water-miscible solvents, e.g. lower alcohols (methanol, ethanol, isopropanol and others~, lower ketones (acetone and others), polar ethers (tetrahydrofuran, dioxane, 1,2-dimethoxyethane and othersJ and to obtain in this way crystallized products which can be easily isolated and readily purified. It proved to be especially advantageous to add the desired base as early as during the complex formation to the reaction mixture and thereby to save a process step.
In case the acidic complex compounds contain several free acidic groups, it is frequently e~pedient to produce neu~ral mixed salts containing inorganic as well as organic cations as the counterions.
This can be done, for example, by reacting the complex forming ligand in an aqueous suspension or solution with the oxide or salt of the element yielding the central ion, and with half the amount of an organic base required for neutralization; isolating the thus-formed complex salt; purifying same if desired; and then combining, for complete neutraliza-tion, with the needed amount of inorganic base. The sequence of addition of the bases can also be reversed.
Another possibility of obtaining neutral complex compounds resides in converting the remaining acid groups in the complex entirely or partially into esters or amides, for example. This can be done by subsequent reaction at the finished complex (e.g. by exhaustive reaction of the free carboxy groups with dimethyl sulfate) : -. ~ ' ' , . .: .

. , : . ~ .' - ' ZO;~i~Z~2 The conjugates of antLbody and complex are dialyz~d, prlor to ln vivo u3e, a~ter inctlb~tlon with .
a weak complexing agent, 5Ucll a~, fer example, ~odium ~itrate, sodium ethylenediaminete~raacetic acld, in order to remove weakly bound metal ato~s.
Tlle pharmaceutl~al agents of this lnv~ntion are likewise produced in a manner known per ~e b~
suspendinq or dissolving the ~omplex compounds of ~th~s invcntlon -~ optionally combined wlth the addi~lves cus~omary in galenic pharma~y -- in an aqueous medium and then optionally sterilizing the suspenslon or solution. Suitable additivQç are, ~or example, physiologi~lly accep~able buffQrs (such as, or lhstance, tromethamin~, addition~ of complexi~g agents (e.g.J~iet~ylenetriam1nepentaacetic acid1 or --if required -- electrolytes, e.g.~sodlum chlor~de o~ --if nece~sary -- antioxldants, su~h as ascorbic acid, or example.
~f suspenslons or solutions o the agents of this invention in water or physiological saline solution are desirable for enteral adminis~ration or other purposes, the~r ~re mixed with one or several of ~he aux~llary agents te.g.~ ~etbyl~ellulose, lactose, manni~ol) and/or tensides ~e.g., 1eaithins, TWEEN~, Z5 MYRJ) and/or flavoring agents ~o impro~e taa~ t~.g., e~hereal oils), a~ cus~mary in galen-~ pharmaey.
In prlnciple, lt ls also possible to produce the pharmaceutical agents of thls lnv~n~ion without isolating the complex ~alt8. In any even~, special ~are must ~e taken to effect chelate formation so th~t the 8alts and ~alt ~olu~ions aacordlng to this ~' .

, . , . ~
.
. .
.

' ' ' ' ' ' -, .:

20342~42 invention ~re practically devoid of uncomplexed, to~ically active metal ions.
This can be ensured, for example, with the aid of dye indicators, such as xylenol orange, by control ~itrations during the manufacturing process.
'l'herefore, the invention also concerns processes for the production of the complex compounds and their salts.
final safety measure resides in purifying the isolated complex salt.
The pharmaceutical agents of this invention preferably contain 0.1 ~mol- 1 mol/l of the complex salt and are normally made into doses in amounts of 0. 1 IJIllOl - S mmol/kg. They are intended for enteral and parenteral administration. The complex compounds according to this invention are utilized tl) for NMR and X-ray diagnostics in the form of their complexes with the ions of the elements with atomic numbers 21-29, 42, 44 and 57-83;
(2) for radiodiagnostics and radiotherapy in the form of their complexes with the radioisotopes of the elements with atomic numbers 27, 29, 31, 32, 37-39, 43, 49, 62, 64, 70, 75 and 77.
The agents of this invention meet the variegated requirements for being suitable as contrast media for nuclear spin tomography. Thus, they are excellently suited for improving the informative content of the image obtained with the aid of the NMR tomograph upon oral or parenteral administration, by increasing the signal intensity. Furthermore, they exhibit the high efficacy necessary to introduce into the body a minimum amount of burdening foreign substances, and ' -' -- 38 _ ~ ~34~2 th~y show ~he good compa~ib~lit~ ~equlred for !`
maintaining ~he nonlnvaslYe ~hara~ter of ~he examina-tio~
The good water ~olubility and lo~ osmolality of the compou~lds of tllis invention ~ake i~ possible to ~repare hi~hly concen~xated solutions, ~hus maintain-ing the volume load on the circulation within tolerable limits and compensating for dilution by bo~y 1uid, i.e., NM~ diagno~tic aid~ ~hould exhib~t 100 - 1,000 ~im~s tS~e watex solubility of agents for NMn spectroscopy.
Furthermor~, the ag~nts~o this invention exhibit not ~only a high in vitro ~tability but also a surprisingly high ~tability in vlvo so ~hat release or ex~hange o~
the ions -- actually toxic -- not bound in a co~alent fashion in the Gomplexes takes place only extremely gradually within the time per~od durin~ which the novel contrast ~edia are again completely elimina~ed.
~ n g~n~ral, the agents of thls invention are used, for NMR diaqnos~ic aids, in dose6 amounting 20 to O.OaOl - $ mmollkg~ preferably O.OOS ~ 0.5 mmol/kg.
De~ails of u~e are discussed, for example, ~n H.J.
Weinm~nn et al., Am. J. of Qoehtgenology 142 ~ G19 ~1984) .
Espacially low doses (b~low 1 ~g/kg body w~ight) of organ-specific NMR diagnostic aids are usabl~
or exan~ple, fox the detection of tumors and of cardiac inarc~ion.
Further~ore, th~ complex compounds ~o~dln~
to this invqntion c:an be employqd with advant~gq as suscep~lbility re~ent~ and a~ s~llf~ reaqents for in vivo NMR spe~troscopy.

: .

2034Zf`~2 The agent~ of this invention, ba~ed on th-ir ~avor~blQ radioactive propert1~ and good st~bllity of the complex compound~ contained therein, are also suited as radiodiagno~ic agen~s. Detail~ of their usa~e ~nd dos~ge are descri.bed, for example, in "~adio~racers for Medical ~ppli~ations", ~R~ Pr~ss, Boca Raton, Florida.
~ nother imaging method with radioisotopes is the positron cmission tomography, u~in~ posi~ron-10 emitting iso~opes, 8uch a~, for example, 43Sc, 44Sc~
52~e, 55Co and 68Ga ~Heiss, W.D.; Phelps, M.E.:
.Positron Emission Tomog~aphy of ~rain, Springer publishers, Berlin, Heidelb~r~, New York l9a3).
The compounds of thl~ invention can al50 b~
utili~ed in radioimmuno- or radi~tion ~herapy. This p~ocess differs from the corresponding di~gnostics only in t21e quanti~y and type of isotope employed.
The obje~tive herein 1~ the d~5truction o tumor c~lls ~y high-energy shortwave radiati~n wlth ~ minimum range.
Suitable ~-emi~ing ions are, for exampl~ 46Sc, 47Sc, 48Sc, 72Ga, 7~Ga and 90Y. Suitable ~-emittlng ions ex-hibiting short half-life pexiods are, for example, llBi, Bi, 138i and ~14Bl, wherein 1 Bi is pre~err~d~ A sui~a~le ion emitting photons and 25 ~lectrons is 15~Gd which can be obtained from 157Gd by neu~ron capture~
~f ~he a~ent of this inventlon i~ intended for use in th~ ve~sion of radiation therapy proposad by R.L. Mill~ et ~1. [Nature~ vol~ 33fi : 787 ~1988)1, then the c~ntral ion ~ust bo derived from a M~ssbauer isotope, such as, for example, 57Fe or lSlEu, Z0;;~4'~2 In the in vivo administration of the therapeutic agents according to this invention, they can be given together with a suitable carrier, e.g.
serum or physiological sodium chloride solution and together with another protein, such as, for example, human serum albumin. The dosage herein is dependent on the type of cellular disorder, the metal ion used, and the type of imaging method, e.g. brachytherapy.
The therapeutic media of this invention are administered parenterally, preferably intravenously.
Details of usage of radiotherapeutic agents are discussed, for example, in R.W. Kozak et al., TIBTEC, October 1986, 262.
The agents of this invention are excellently suited as X-ray contrast media; in this connection, it is to be especially emphasized that they reveal no indication of anaphylaxis-type reactions, known from ~;
iodine-containing contrast medi~, in biochemical-pharmacological studies. They are particularly valuable, on account of the favorable absorption properties in regions of higher tube voltages, for digital subtraction techniques.
In general, the agents of this invention are utilized, for administration as X-ray contrast media, ~
25 analogously to, for example, meglumine diatrizoate, -in doses amounting to 0.1 - 5 mmol/kg, preferably 0.25 - 1 mmol/kg.
Details of utilization of X-ray contrast media are discussed, for example, in Barke, "Rontgenkontrast-mittel" [X-Ray Contrast Medial, G. Thieme, Leipzig ~1970) and P. Thurn, E. ~cheler, "Ein~hrung in die Rontgendiagnostik" EIntroduction to X-Ray Diagnostics], G. Thieme, Stuttgart, New York (1977).
, , .

.
~' -~ ' '''. ' : ' X0;~4;~`2 In summa~i~sl, ~ho ~ynthesi~ ha~ b~naco~pl ~ ~hed ~æ novel complexing ao~pound~, metal co~plexe~ and ~ethal ao~plex ~alt~, opening up n~w posslbilitles ln ~lagnostic and therapeu~ic ~edicine.
Thi~ development appears to be de~irahle, above all in light of t~le ~volutioll of novel imaging methods in medical diagno~tics.
Wlthout further elaboration, it is believed that one skilled in the art can, usin~ the preaeding de~cription, utilize ~he pres~nt invention to its fullest extent. The ~ollowing preferred speaific ambod~ments ar~, therefore, to bs con~trued as merely illustratlve, and not limitative of the rem~lnder of the dl~clo~ure in any way whatsoever.
In the foregoing and in the followin~ example~, all temperatures are set forth uncorrected in deqree~
Celsius and unless otherwise indicated, all parts and percentages are by weight.
The entire disclosures of all applications, patents and publicat~ons, i~ any, cited above an~ below, and of corresponding applic~tion German P 40 01 ~55.2, filed January 18, 1990, are hereby lnoorporated by reference.

' ~

.
., ,"., '-- ~2 -ZV3~2~2 xamT~le 1 (a) 3,6,9-Tris[dihydro-2(3H)-furanon-3-yl]-3, 6, 9, 15-tetraazabicyclo[9.3.1~pentadeca-1(15),11,13-triene ___________________________________________________ 40 g (242.38 millimoles) of ~-bromo- ~-butyro-lactone is added to 10 g (48.48 mmol) of 3,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene, 33.5 q (242.38 mmol) of potassium carbonate and 805 mg (4.85 mmol) of potassium iodide in 200 ml of acetonitrile. This mixture is heated under reflux for 48 hours, evaporated to dryness under vacuum, and the residue is taken up in ~00 ml of methylene chloride and extracted three times with 150 ml of water. The organic phase is dried over magnesium sulfate and evaporated under vacuum. The residue is purified by chromatography on silica gel, (Mobile phase:
methylene chloride/methanol = 15:1) Yield: 7.11 g (32% of theory) of a slightly yellow-colored oil which solidifies when allowed to stand. .
Analysis:
C 60.25 H 6.59 N 12.22 (Calcd.) -C 60.18 H 6.64 N 12.17 (Found) .

(b) Gadolinium Complex of 3,6,9,15 Tetraazabicyclo- -~9.3.1]pentadeca-1(15),11,13-triene-3,6,9-tris-25~a-(2-hydroxyethyl)acetic Acid]
_________________________________ ____________ .
6.7 g (14.61 mmol) of the title compound o~ Ex-ample l~a) is dissolved in 50 ml of deionized water, and the pH is brought to 5.5 by adding l-normal hydro-ohlor ic acid . To thi s n~ixture is added 2 . 65 g ;

~03~2~tZ

(7.~ mol) o~ c~aclolinium oxide and the mixture is reEluxed Lor 3 l~ours. The cooled solution is stirred for one l~our with respectively 10 ml of acidic ion exchanger (IR 120) and 10 ml of alkaline ion exchanger (IR~ 410). The mixture is filtered off from the ex-chan~er, ancl the filtrate is boiled for one hour with active carbon. After filtration and freeze-drying, 9.25 9 (95% of theory) of an amorphous, colorless powder is obtained (containing 8.3% of water yer analysis).

Analysis (corrected for water):
C 41.43 El 4.99 N 8.40 Gd 23.58 (Calcd.) C 41.35 El 5.09 N 8.34 Gd 23.50 (Found) (c) Europium Complex of 3,6,9,15-Tetraazabicyclo-[9.3.1]pentadeca-1(15),11,13-triene-3,6,9-tris[~-(2-hydroxyethyl)acetic Acid]
_____________________________________________ Analogously, the corresponding Europium complex is obtained with 151Eu203.

Analysis(corrected for water):
20 C 41.76 H 5.03 N 8.47 Eu 22.97 (Calcd.) C 41.68 3 5.12 N 8.39 ~u 2.88 ~ound) ' ~ .

. .

.. ; .

" .:
-~ : .,.. ~ .: ~
,, ` ; ': ' ; ~
:

Z034Z~2 (~) 3,6,9,15-'retraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene-3,6,9-tris~ enzyloxy-methyl)acetic Acid]
_______________ ___________________________ 10 g (48.48 mmol) of 3,6,9,15-tetraazabicyclo-~9.3.1]pentadeca-1(l5),11,13-triene and 114.71 g (48~.8 mmol) of the sodium salt of 2-chloro-3-benzyloxy-propionic acid in 200 ml of water are heated to 70 C
for 48 hours. The solution is diluted with 400 ml of water and 300 ml of 2N hydrochloric acid is added thereto. The mixture is extracted 5 times with re-spectively 200 ml of m~thylene chloride. The aqueous phase is evaporated under vacuum. The residue is dissolved in 300 ml of ethanol and filtered off from the sodium chloride. Then the mixture is evaporated under vacuum and the remaining oil is chromatographed on silica gel Imobile phase: ethanol/water = 20:1~.
The main fractions are evaporated under vacuum and dis-solved in 50 ml of 5~ strength hydrochloric acid. The solution is passed over a column, filled with "Reillex" (= poly-4-vinylpyridine), and the product is eluted with a mixture of water/methanol 3:1.
After evaporation of the main fractions, 12.93 g 136% of theory) of a strongly hygroscopic solid is obtained (9.1% water per analysis).

Analysis (corrected for water):
C 66.47 H 6.53 N 7.56 (Calcd.) C 66.38 H 6.60 N 7.48 (Found) ' .

. .

' - ~5 -;~()342~2 (1~) 3,~,9,15-T~raazabi~yC10~9.3.1]pentadecil-~ ),11,13-triene-3,6,9-t~-isl~-(hydroxy-methyl)acetic Acid]
______________________________ __________ 12 h g (17.01 mmol) of the title compound of F.xample 21~) is di~solved in a mixture of 200 ml of meth~nol/100 ~1 of water, and 4 ~ of palladium ca~alyst is added (10~ Pd on active carbon). The mixt~re is hydrogetlated for 5 hours ~t 50 C, ~ ered off from the catalyst, and evapo~a~e~ ~nder vacuum.
Yield: 7.84 g (98~ of tlleory) of a vitreou~ solid (6.9% wa~er per analysis).
~, Analysis (corrected for water):
C 51.06 H 6.43 N 11.91 (Calcd.) C 50.97 H 6.51 N 11.81 IFound) lS (c) Gadolinium Complex of 3,6,g,1~-Tetraazabicy~lo-9.3.11pentadeca-1(15),11,13-tri~ne-3,G,~-tris-la-~hydroxymethyl)~cet~c ~cidl _______________________ ____________ ________ 7.5 g (15.94 mmol) of thc title ~ompound of Example 2(b) is dissolved in S0 Inl of deionized water, and 2 . 89 g (7 . 97 n~nol~ of gado~inium oxide is added.
The mixture is heate~ for 3 hours at 90 ~ The cooled solution is stirred for onc hou~ at room temperature wi~h respectively 2 ml of ~cidic ion exchangcr (IR 120) and 2 ml of ~1kflline exchanger IIRA 410), ~1tqred off from the ~xchanyer, and tho fil~ate i~ briefly boil~d with active carhon. Af~er filtr~tion and ~e~e-dryinq, 9.5~ ~ ~9~ of theory) of ~ oolorless, amclrphous powder ls oOtained ~11.1~ water per analyslt ) .

.
, : ' :
, .
': ~` .
.

3~2q`2 ~nalysi~ (corr~ct~d for water):
C 38.45 ll 4.36 N 8.97 Gd 2~.17 (Calcd.) C 38.37 1l 4.43 N 8.89 Gd 25.06 (Found) Example 3 (a) 3,6,9,15-Tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene-3,6,9-tris[~-(1,2-O-isopropyl-idene-1,2-dihydroxyethyl)acetic ~cid Ethyl Ester]
_______----A mixture of 15 g (72.71 mmol) of 3,6,9,15-tetra-azabicyclo[9.3.1]pentadeca-1(15),11,13-triene, 156.38 g ~426.28 mmol) of 3,4-O-isopropylidene-2-(p-tolylsulfonyl)-3,4-dihydroxybutyric acid ethyl -ester, 60.3 g (436.28 mmol) of potassium carbonate, ~-and 2.41 g (14.54 mmol) of potassium iodide in 400 ml of ace*onitrile is heated under reflux for 48 hours.
The mixture is evaporated under vacuum and the residue taken up in 500 ml of methylene chloride. The mixture is extracted 3 times with 200 ml of water and the organic phase is dried over magnesium sulfate. After evaporation, the remaining oil is chromatographed on silica gel (mobile phase: methylene chloridethexane/
methanol = 20:4:1).
Yield: 17.24 g (31~ of theory) of a yellow, viscous oil.

Analysis:
C 59.67 H 7.91 N 7.32 (Calcd.) C 59.59 H 7~98 N 7.27 ~Found) .. ~

:, ,~ . ' .

- ~7 -;~034;2~2 (b) 3,G,9,l5-Tctrcl~lzabicyclol9.3.1~pentadeca-1(15),11,13-triene-3,6,9-tris[a-(1,2-dihydroxy-etllyl)acetic Acid~
_______________________________________________ 16.5 g (21.57 mmol) of the title compound of Ex~ple 3(a) is dissolved in 100 ml of ethanol, and 50 ml of 5N sodium hydroxide solution is added thereto.
The mixture is heated under reflux for 10 hours and evaporated under vacuum. The residue is dissolved in 250 ml of methanol and filtered off from the sodium chloride. The filtrate is evaporated under vacuum and the residue purified on an ion exchanger as follows:
The product is dissolved in S0 ml of water, and the solution is introduced into a cation exchange column (IR 120). After flushing with water, the ligand is eluted with 0.5N aqueous ammonia solution. The main fractions are evaporated, taken up in a small amount of water and passed over an ion exchange column (IRA 67).
The product is first washed with water and then eluted with 0.5N formic acid. The mixture is evaporated under vacuum and the residue dissolved in a small quantity of hot methanol. By the gentle addition of acetone and cooling in an ice bath, the title compound is obtained in crystalline form.
Yield: 8.22 g (68% of theory) of a vitreous solid (9.2% water per analysis).
.
Analysis (corrected for water):
C49.28 H 6.47 N 9.99 (Calcd.) C49.17 H 6.56 N 9.88 (Found) . . :
.

:. : ., ~ . ' ~ .

:' ~

~0342~2 (c) G~{lolinialm Complcx of 3,6,9,15-Tetraazabicyclo-[9.3.1]pen~adeca-1(15),11,13-triene-3,6,9-tris-[~-(1,2-dihydroxyethyl)acetic Acid]
_____________________________~_________________ 8 g (14.27 mmol) of the title compound of Exampl~ 3(b) i~ dissolved in 60 ml of deionized ~ater, and 2.58 g (7.135 mmol) of gadolinium oxide is added.
The mixture is heated for 3 hours at 90 C. The -.
cooled solution is stirred for one hour at room tem-perature with respectively 2 ml of acidic ion exchanger 10 ~IR 120) and 2 ml of alkaline exchanger (IR~ 410). The mixture is removed from the exchanger by filtration and the filtrate boiled with active carbon. After fil~ra-tion and freeze-drying, 9.89 g (97% of theory) of a colorless, amorphous powder is obtained (containing 7.3~ water per analysis).

Analysis (corrected for water):
C 38.65 H 4.65 N 7.84 Gd 22.00 (Calcd.) C 38.54 H 4.74 N 7.78 Gd 21.92 (Found) Example 4 20 (a) 2,2',6,6'-Tetra(hydroxymethyl)-4,4'-bipyridine ____________________________________ ___ _____ 50 g (128.77 mmol) of 2,2',6,6'-tetra(methoxy-carbonyl)-4,4'-bipyridine is dissolved in a mixture of 400 ml of dioxanel400 ml of water and, in portions, 48.71 g (1.28 mol) of sodium borohydride is added thereto. The mixture is stirred overnight at xoom temperature. The solution is acidified with 5N
hydrochloric acid and evaporated to dryness. The residue is suspended in 1 liter of lN sodium hydroxide , :.

.

- 4~ -203~2~Z

solutiol~ all~l c~r~lcted thr~e times with 250 ml of clllo~o~orm. The organic phases are dried over magnesium sulEate and evaporated under vacuum. The residue is recrystal]ized from ethanol/ether.
Yield: 29.53 g (83~ of theory) of colorless crystals.

Analysis:
C 60.86 ~1 5.84 N 10.14 (Calcd.) C 60.77 ll 5.93 N 10.06 (Found) (b) 2,2',6,6'-Tetra(chloromethyl)-4,4'~bipyridine _____________________________________________ 29 g (104.96 mmol) of the title compound of Example 4(a) is heated under reflux for 5 hours in 250 g (2.1 mol) of thionyl chloride. The mixture is evaporated to dryness and the residue taken up in 200 ml of concentrated soda solution. The mixture is extracted twice with 150 ml of methylene chloride.
The organic phase is dried over magnesium sulfate and ~-evaporated under vacuum. The residue is crystallized from ether/hexane.
Yield: 35.54 g (94% of theory) of colorless crystals.

Analysis:
C 48.03 H 3.45 N 8.08 Gd 40.51 (Calcd.) C 48.10 H 3.40 N 7.96 Gd 40.59 (Found) ' ~ -, ~-- . . ~.

.:'~
. ~ . ... ...
' : . -.:~ - .

, X03~2~L~2 (c) 13, 13 ' -13is 13,6,9-tris(p-tolylsulfonyl~-3,6,9,15-t:etraazabicyclol9.3.1~pentadeca-1(15),11,13-triene]
_________________________________~______________ ~ t 100 C, a solution of 34 g (97.12 mmol) of the title compound of Example 4(b) (dissolved in 700 ml of dimetllylformamide) is addded dropwise within 4 hours to 118.43 g (194.25 mmol) of N,N',N''-tris(p-tolyl-sulfonyl)diethylenetriamine-N,N''-disodium salt in 1600 ml of dimethylformamide. The mixture is stirred overnight at 100 C. Two liters of water are added dropwise to the hot solution, and the latter is allowed to cool down to 0 C. The precipitate is suctioned off and washed with water. After drying under vacuum (60 C), the product is recrystallized from aceto-nitrile, thus obtaining 79.13 g (61~ of theory) of a cream-colored powder.

Analysis:
C 57.55 H 5.28 N 8.39 S 14.40 (Calcd.) C 57.47 H 5.35 N 8.13 S 14.32 (Found) (d) 13,13'-Bis[3,6,9,15-tetraazabicyclo[9.3.11penta-deca-1(15),11,13-trienel Octahydrosulfate ________ ______ _________ ______________________ 79 g (59.15 mmol) of the title compound of Example 4(c) is introduced into 270 ml of concentrated sulfuric acid and stirred for 48 hours at 100 C, then cooled to 0 C, and 1.35 1 of absolute ether is added dropwise. The precipitate is suctioned off and extracted by stirring in 500 ml of methanol. After the product has been filtered off and dried under vacuum, 65.74 g ~93% of theory) of a solid is obtained which deliquesces in the air.

., '' . ' ' , ' ........ :'- ' .

,. ~, ~ . .

: .

Z~)3~ Z
ula I y s i s ~ 22.11 ll ~.22 N 9.38 S 21.46 (Calcd.
C 22.04 1l 4.33 N 9.29 S 21.38 (Found) (e) 13,l3' -Bis[ 3,6,9,15-tetraazabicyclo[9.3.1]penta-cleca-~(15),1],13-triene]
_________________________________________.______ 65.5 g t5q.80 mmol) of the title compound of Example 4(d) ls dissolved in 100 ml of water and the pH is adjusted to 13 with 32% strength sodium hydroxide solution. The mixture is extracted three times with 250 ml of hot toluene. The combined toluene phases are heated under reflux for one hour with 20 g of finely pulveriæed sodium hydroxide. ~he mixture is filtered and the filtrate evaporated to dryness.
Yield: 21.6 g (96~ of theory) of a solid having a slightly yellow color.

Analysis:
C 64.36 H 8.35 N 27.29 (Calcd.) C 64.27 H 8.44 N 27.22 (Found) (f) 13,13'-sis[3,6,9,15-tetraazabicyclo[9.3.1]penta-deca-1(15),11,13-triene-3,6,9-trist~-(benzyloxy-methyl)acetic Acidl]
_______________ ________________________________ 21.5 g (52.37 mmol) of the title compound of Example 4(e) and 247.8 g (1.05 mol) of the sodium salt of Z-chloro-3-benzyloxypropionic acid in 400 ml of 25~ water are heated for 48 hours to 70 C. The solution is diluted with 800 ml of water and combined with 600 ml of 2N hydrochloric acid, then extracted 5 times with respectively 300 ml of methylene chloride, and the aqueous phase is evaporated under vacuum. The . ~, .. . . . . . .

.

, ~ , ~03~Z~iZ
residlle is ~lissolved in 500 ml of ethanol and filtered ofE Erom tlle sodium chloride. The mixture is evaporat-ed under vacuum and the residue chromatographed on silica gel (mobile phase: ethanol/water = 20:1).
The main fractions are evaporated under vacuum and dis-solved in 100 ml oE 5% strength hydrochloric acid.
The solution is passed over a column filled with "Reillex" (= poly-4-vinylpyridine), and the product is eluted with a mixture of water/methanol 2:1. After evaporation of the main fractions, 20.92 g (27% of theory) of a strongly hygroscopic solid is obtained (8.1% water per analysis).

Analysis (corrected for water):
C 66.56 ll 6.40 N 7.57 (Calcd.) lS C 66.47 H 6.51 N 7.48 (Found) (g) 13,13'-BisL3,6,9,15-tetraaZabicyclo[9.3.1]penta-deca-1(15),11,13-triene-3,6,9-tris[~-(hydroxy-methyl)acetic Acidl]
_______________________________________________ 20.5 g (~3.85 mmol) of the title compound of Example 4(f) is dissolved in a mixture of 300 ml of methanol/150 ml of water, and 7 g of palladium catalyst (10% Pd on active carbon) is added thereto. The mix-ture is hydrogenated for 5 hours at 50 C. The product is removed from the catalyst by filtration and evaporat-ed under vacuum.Yield: 12.62 g (97% of theory) of a vitreous solid (8.5% water per analysis).
Analysis ~corrected for water):
C 51.17 H 6.23 N 11.93 (Calcd.) 30 C Sl.07 H 6.31 N 11.87 (Found) . . - . :
.. , ~. , ' ' '.
~ ' ` ';.

. .

203~2~2 a-lo~inium Complex of 13,J3'-L~is [ 3,6,9,15-tetra-aza~icyclol9.3.1]pentadeca-1(15),11,13-triene-3,6,9-tris[~-(hydroxymethyl)acetic Acidl]
________________________________________________ 12 g (12.78 mmol) of the title compound of Example 4(g) is dissolved in 80 ml of deionized water, and 4.63 g (12.78 ~nol) of gadolinium oxide is added.
The mixture is heated for 3 hours at 90 C. The drawn-off solution is stirred for one hour at room temperature with respectively S ml of acidic ion exchanger ~IR 120) 1~ and 5 ml of alkaline exchanger (IRA 410). The product is filtered off from the exchanger, and the filtrate is briefly boiled with active carbon. ~fter filtration and freeze-drying, 15.3 g (96~ of theory) of a colorless, amorphous powder is obtained (containing 9.3% of water per analysis).

Analysis (corrected for water):
C 38.52 11 4.20 N 8.98 Gd 25.21 (Calcd.) C 38.46 H 4.28 N 8.91 Gd 25.14 (Found) Example 5 C (a) Trans-5-(p-tolylsulfonyl)amino-6-(p-tolyl-sulfonyloxy)-2,2-dimethyl-1,3-dioxepane ____________ ______________________ ______ Under agitation, 295.67 g of p-toluenesulfo-chloride is added in portions at -5 to 0 C to a solution of 100 g of trans-6-amino-2,3-dimethyl-1,3-dioxepan-5-ol in 903 ml of p~ridine. The mixture is allowed to stand for 72 hours at +4 C and then stirred into 10 1 of ice water. After the precipitate has been suctioned off and washed with water, the residue is dried in a drying cabinet at 50 C and 200 torr for 48 hours. For purposes of purification, the crude : , , ~ . ..

' -' ,~ .

~034XI~Z

produc~ is r~crystallized ~rom 5 1 of dioxane, thus ob-taining 196 9 of the title compound as a white powder, mp 200-202 C.

(b) Monosodium Salt of N-[2-(N-Tolylsulfonylamino)-ethyl]-p-tolylsulfonamide _______________________________________________ 150 g of N-[2-(N-tolylsulfonylamino)ethyl]-p-tolylsulfonylamide is suspended in 1.25 1 of ethanol, heated under reflux, and a solution of 10.3 g of sodium in 300 ml of ethanol is added dropwise thereto, thus forminq a solution. During cooling, the title compound is precipitated, suctioned of, the precipitate washed with ethanol and dried at 50 C and 200 torr. Yield:
119 g of the title compound as a white powder.

(c) cis-2,2-Dimethyl-5-[N-(p-tolylsulfonyl~amino]-6-[N-(p-tolylsulfonyl)-N-(N'-2-p-tolylsulfonyl-aminomethyl)1-1,3-dioxepane ________ _______________________________ _______ 116 g of the monosodium salt of Example 5(b) is suspended in 2.66 1 of dimethylformamide. At 100 C, a solution of 141 g of trans-5-(p-tolylsulfonyl)amino-6-(p-tolylsulfonyloxy)-2,2-dimethyl-1,3-dioxepane in I.5 1 of dimethylformamide is added dropwise thereto and the mixture is stirred for 5 hours at a bath temperature of 120 C. The reaction solution is then concentrated under vacuum to 1 liter and diluted with 10 1 of ice water, suctioned off, the precipitate is washed with water and dried at 50 C and 200 torr, and the product is 182 g of the crude title compound.
For purification, the product is extracted by boiling wïth 1.85 1 of ethanol. After suctioning off and drying, 125 g of the title compound is obtained as a white powder, mp 190-194 C.
' ' , " . ~
' Z03~
(d) I)i;o~iulli Salt of cis-2,2-Dime~hyl-5-[N-(p-tolyl-sulfonyl)amino]-6-[N-(p-tolylsulfonyl)-~-(N'-2-p-tolyLsulfonylaminomethyl)l-1,3-dioxepane ________________________________________________ 87.8 g of the compound obtained according to I~:xample 5(c) is suspended in 410 ml of ethanol, heated to boiling, and a solution of 6.67 g of sodium in 200 ml of ethanol is added dropwise thereto. The mixture is cooled in an ice bath, combined with 450 ml of ether, and suctioned off from the precipitate, which latter 10 is dried at 80 C and 200 torr, thus producing 91 g of the title compound as a white powder.

~e) Acetonide of 4,5-sis(hydroxymethyl)-3,6,9-tri-tosylsulfonyl-3,5,9,15-tetraazabicyclo[9.3.11penta-deca-1(15),11,13-triene _ _ _ _ _ _ :
7~.75 g of the disodium salt of Example 5(d) is dissolved in 880 ml of dimethylformamide, heated to 100 C, and a solution of 19.53 g of bis(2,6-chloro-methyl~pyridine in 360 ml of dimethylformamide is added dropwise thereto; the mxiture is heated for 5 hours to 20 120 C and concentrated under vacuum to 300 ml. The solution is stirred into 5 1 of ice water, the precip-itate is suctioned off, washed with water, and dried.
The crude product is recrystallized from 700 ml of dioxane, thus obtaining 45 g of the title compound as 25 a white powder, mp 244-250 C.

- ,6 '~o342~f2 (f) ~c~oni~ ol ~,5-~is(l~ydroxymethyl)-3,6,9,15-tetra-a~abicyclo[9.3.1]pelltadeca-1(15),11,13-triene ___________________________________________________ ~ suspension of 20 9 of the compound obtained according to Example 5(e) in 140 ml of tetrahydrofuran is added to 260 ml oE liquid ammonia; the mixture is stirred in a refrigerating bath at -50 C, and a total of 14.4 g of sodium is introduced in portions. The mixture is stirred for another 5 hours at -60 C, then the refrigerating bath is removed, and S0 ml of ethanol is added dropwise. The ammonia is allowed to evaporate, thc mixture is evaporated to dryness under vacuum, and the residue is purified by chromatography on silica gel. Elution with chloroform/ethanol/concentrated ammonia solution l3/1/0.5) yields 5.30 g of the title compound as an oil.

Analysis:
C 62.72 ~ 8.55 N 18.29 (Calcd.) C 62.51 H 8.41 N 18.45 (Found) (g) Acetonide of 4,5-Bis(hydroxymethyl)-3,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene-3,6,9-tris~acetic Acid tert-Butyl Ester) _______________________ _______________________ 5.51 g of anhydrous sodium carbonate and 10.2 g of bromoacetic acid tert-butyl ester are added to a solution of 4 g of the compound produced according to 25 Example 5~) in 100 ml of tetrahydrofuran and 10 ml of water and the mixture is stirred for S hours at 50 C. The mixture is filtered, evaporated under vacuum, and the oily residue is stirred with 50 ml o -hexane and decanted. The residue i9 purified by ' ' ', ' .... ~

~034Zl$2 cllrom~lto~31-aL~Ily on 100 9 of silic~ gel with dichloro-~netl)~ne (1-10~ ethallol), thus obtaining 5.7 g of the title compound as a light-yellow oil.

(h) 4,5-Bis(hydroxymethyl)-3,6,9,15-tetraaza-bicyclo~9.3.1]pentadeca-1(15),11,13-triene-3,6,9-~ris-acetic Acid ___________________________________________ A mixture of 5.3 g of the ester prepared accord-ing to Example 5(g) and 50 ml of trifluoroacetic acid is stirred for 3 hours at 50 C. Then 10 ml of water is added, -the mixture is stirred for another 2 hours at 50 C, and thereafter evaporated to dryness under vacuum. The residue is dissolved in 20 ml of water, and the solution is allowed to pass through a column with 100 ml of "Reillex" (poly-4-vinylpyridine), eluted with 100 ml of water, and the eluate is evap-orated under vacuum, thus obtaining an amorphous powder which still contains 8.5~ water.
Yield: 2.90 g.

Analysis:
20 C 51.81 H 6.41 N 12.72 (Calcd.) C 51.63 H 6.70 N 12.51 (Found) (i) Gadolinium Complex of 4,5-Bis(hydroxymethyl)-3,6,9,15-tetraazabicyclo(9.3.1]pentadeca-1(15),11,13-triene-3,6,9-tris-acetic Acid ____________________ ________________________ 2 g (water content 8.5%, corresponding to 1.83 g = 4.29 mmol) of 4,5-bis~hydroxymethyl) 3,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene-3,6,9-tris-acetic acid (production see Example 5h) and 778 mg of gadolinium oxide are stirred for 5 hours ?

' ~-:
, Z03A2~2 wi~h 50 ml o wclter at 90 C. Af~er cooling, the mix-t:ure is ~tirred in succession with respectively 10 ml of anion exchanger IR~ 410 and cation exchanger IRC 50, Eiltered, and the solution is sub~ected to freeze-drying, thus obtaining 2.35 g of the title compoundas a loose wlli~e powder, water content, according to K.-Fiscller titration: 7.3%.

Analysis (after correction for water content):
Clsll2sGdN48 10C 38.38 H 4.24 N 9.42 Gd 26.44 (Calcd.) C 38.51 El 4.31 N 9.36 Gd 26.19 (Found) Example 6 (a) Acetonide of 4,5-Bis(hydroxymethyl)-3,6,9-tris-[dihydro-2-(3H)-furanon-3-yl]-3,6,9,15-tetraaza-15bicyclo(9.3.1]pentadeca-1(15~,11,13-triene ________________________________________________ A solution of 5 g (16.34 mmol) of acetonide of 4,5-bis(hydroxymethyl)-3,6,9,15-tetraazabicyclo[9.3.1]-pentadeca-l (l5! ,11,13-triene in 100 ml of acetonitrile is combined with 12 g of potassium carbonate, 260 mg of potassium iodide and 13.50 g of a-bromo- ~-butyro-lactone and heated for 48 hours to boiling. The mix-ture is then evaporated under vacuum, the residue is dissolved in methylene chloride, shaken several times with water, the organic phase is dried over sodium sulfate, and evaporated to dryness. The oily residue ls chromatographed with methylene chloridetmethanol (15:1) on 150 g of silica gel, thus obtaining 5.3 g of the title compound as a light-yellow, viscous oil.
;

~ - , .
' , :, .

_ 59 ~
X03~2~$2 ~b) 4,5-sis(llydroxymethyl)-3~6~9~l5-tetraaza-bicyclol9.3.1]pentadeca-1(15),11,13-triene-3,6,9-tris~-(2-hydroxyethyl)]acetic Acid ___________________________________________ 5 g of the compound produced according to Example 6(a) is dissolved in S0 ml of water and adjusted to a p~l of 2 by addition of hydrochloric acid. The mixture is heated for 5 hours under reflux, cooled to room temperature, and the solution is allowed to pass over a column with 10 g of "Reillex"
(poly-4-vinylpyridine). The column is rinsed with 20 ml of water and the combined eluates are subjected to freeze-drying, thus obtaining 4.05 g of the title compound as a loose powder with a water content of 7.2~.

Analysis (after correction for the water content):
c2sH4ON4oll C 52.44 H 7.04 N 9.78 (Calcd.) C 52.61 H 7.33 N 9.62 (Found) (c) Gadolinium Complex of 4,5-Bis(hydroxymethyl)-3,6,9,15-tetraazabicyclol9.3.1]pentadeca-1(15),11,13-triene-3,6,9-tris[a-(2-hydroxy-ethyl)]acetic Acid _____________________________________________ 1.50 g (2.68 mmol) of 4,5-bis(hydroxymethyl)-3,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene-3,6,9-trisla-(2-hydroxyethyl)]acetic acid is stirred in 25 ml of water with 487 mg of gadolinium oxide for 4 hours at 90 C. After cooling, the solu-tion is stirred in succession with 5 ml of anion exchanger IRA 410 and 5 ml of cation exchanger IRC 50, filtered, and subjected to freeze-drying, ' ' ` .
~ -X034Z~2 t:llUs obtainil~g 1.69 g of the title compound as a white powder with cl wat~î content of ~.3~.

~nalysis (after correction ~or the water content):
~25ll37GdN~ioll C 41.3] Il 5.13 N 7.71 Gd 21.63 (Calcd.) C 41.07 11 5.33 N 7.61 Gd 21.89 (Found) Example_7 (a) 4-Hydroxymethyl-3,6,9-tritosyl-3,6,9,15-tetraaza-bicyclo[9.3.1]pentadeca-1(15),11,13-triene _________________________________________________ A solution of 59.57 g of 3-aza-1-hydroxymethyl-1,3,5-tritosylpentadiamine (preparation see Inter- ~A
national Patent Application PCT/DE 88/00200, WO
88/08422, page 45) in 500 ml of dimethylformamide is combined, in portions, with 9.60 g of a 50~ strength suspension of sodium hydride in mineral oil, and the mixture is heated for one hour to 80 C. To this solution is added dropwise 17.61 g of 2,6-bis(chloro-methyl)pyridine dissolved in 150 ml of dimethyl-formamide, and the mixture is heated for 6 hours to 110 C. The mixture is concentrated under vacuum to about 220 ml, and 1 liter of water is added dropwise.
The precipitate is suctioned off, washed with water, and dried overnight at 50 C and 200 mbar. The crude product is recrystallized from 500 ml of ethanol and yields 45 g of the title compound as a yellow solid.

.~, . . .
:

, . ~: ~ :

- : :

- 6l - ~034~ 2 (b) 4-1lydroxylllethyl-~,6,9,15-te~raazabicyclo[9.3.1]-pelltadeca-1(15),11,13-triene ________________________________________________ ~ 2 g of the compound obtained according to Ex-ample 7(a) is heated Witll 120 ml of concentrated sulfuric acid for 48 hours to 100 C. The mixture is cooled to 0 C, and 350 ml of diethyl ether is added dropwise. The salt of the title compound is thus precipitated. The mixture is suctioned off, the residue is dissolved in 100 ml of water and combined with 40 g of sodium hydroxide and extracted repeatedly with dichloromethane. The combined organic phases are dried over sodium sulfate and evaporated under vacuum, thus obtaining 13.3 g of the title compound as a viscous oil.

(c) 4-Hydroxymethyl-3,6,9,15-tetraazabicyclo[9.3.1]-pentadeca-1~15),11,13-triene-3,6,9-triacetic Acid _________________________________________________ 10 g of the amine prepared according to Ex-ample 7(b) is dissolved in 100 ml of water and combined with 13.21 g of chloroacetic acid. The mixture is stirred for 5 hours at 60 C, maintaining the pH
during this time at 9.0 by addition of lON sodium hydroxide solution. The mixture is cooled to 0 C, combined with 100 ml of ethanol, and acidified with concentrated hydrochloric acid to pH 1. The thus-formed precipitate is suctioned off, dissolved in50 ml of water, and the solution is passed over a column with 30 ml of "Reillex" (poly-4-vinylpyridine), rinsed with 50 ml of water, the eluates are combined and subjected to freeze-drying. Yield: 16.8 9 of - 30 the title compound as an amorphous powder with a water content of 9.3%.

?`

: . , :,, - , .

': : , ' ~
.,, ;

~0342~Z

~llalysis (a~t~r correction for the water content):
C18~l26N4 7 ( 52.68 ll 6.39 N 13.65 (Calcd.) C 52.49 ll 6.54 N 13.81 (Found) S (d) Gado]inium Complex of 4-llydroxymethyl-3,6,9,15-tetraazabicyclo[9.3.1~pentadeca-1(15),11,13-triene-3,6,9-triacetic Acid _______________________________________________ A solution of 2.3 g of 4-hydroxymethyl-3,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene in 10 30 ml of water is combined with 1.015 g of gadolinium oxide and heated for one hour to 90 C. The product is removed from a small amount of unreacted oxide by filtration, and the solution is allowed to run in succession over respectively 10 ml of anion ex- ~-lS changer IRA~410 and cation exchanger IRC 50, rinsed with 30 ml of water, and the combined eluates are subjected to fxeeze-drying. Yield: 3.05 g of the title compound as a powder with a water content of 7.5%.

Analysis (after correction for the water content):
C 38.29 H 4.11 N 9.92 Gd 27.85 (Calcd.) C 38.44 H 4.32 N 9.68 Gd 27.71 (Found) :: ~

. ~ ' :, ~', ' ` ' ~.

- (,3 -Z0;~42~.~
..x .
(a) 4-Benzyloxymethyl-3,6,9,15-tetraa%abicyclo~9.3.1]-pentadeca-1(15),11,13-triene-3,6,9-triacetic Acid __________________________________________________ A solution of 10 g (~- 24.36 mmol) of 4-hydroxy-methyl-3,6,9,15-tetraazabicyclol9.3.1]pentadeca-1(15),11,13-triene-3,6,9-triacetic acid (prepared according to Example 7b) in 150 ml of dimethylformamide is combined with 0.5 g of potassium iodide, 4.17 g of benzyl bromide, and 5 g of sodium carbonate. The mix-ture is heated for 20 hours to 60 C, concentrated under vacuum, combined with 100 ml of water and 300 ml of ethanol, and the pH is set at 2 by addition of concen-trated hydrochloric acid. The mixture is suctioned off from the precipitate, the precipitate is dissolved in 100 ml of water, and the solution is passed over a column with 50 g of "Reillex" (poly-4-vinylpyridine).
The column is washed with 50 ml of water and the com-bined aqueous phases are subjected to freeze-drying, thus obtaining 8 g of the title compound as an amorphous powder.

Analysis:
C 59.99 H 6.44 N 11.19 (Calcd.) C 59.71 H 6.49 N 11.38 (Found) : - , . ,; ~ :
.

~, - ~4 -X0342~-12 (b) (:;~dolilliulll Complex of 4-Benzyloxymethyl-3,6,9,15-tetraa~abicyclo[9.3.1]pentadeca-1(15),11,13-triene-3,6,9-triacetic Acid _________________________________________ 1.~1 g of gadolinium oxide is added to a solution of 5 g of 4-benzyloxymethyl-3,6,9,15-tetraazabicyclo-[9.3.1]pentadeca-l(15),11,13-triene-3,6,9-triacetic acid in 100 ml of water; the mixture is heated for 3 hours to 80-90 C, filtered, and the solution is allowed to pass in succession over columns with re-spectively 15 ml of anion exchanger IRA 410 and cation exchanger IRC 50. The columns are rinsed with 75 ml of water, and the combined aqueous phases are subjected -to freeze-drying, thus obtaining 5.85 g of the title compound as an amorphous powder, water content 7.4%.

Analysis (after correction for water content):
C 45.86 H 4.4~ N 8.56 Gd 24.02 (Calcd.) C 45.69 H 4.71 N 8.72 Gd 23.81 (Found) Example 9 ,.
Gadolinium Complex of 3,6,9-Tetraazabicyclo[9.3.1]-pentadecane-3,6,9-tris[~-(hydroxymethyl)acetic Acid]
__ ___________ ____ ________________________________ 4.5 g (7.2 mmol) of the title compound of Example 2(c) is dissolved in 150 ml of deionized water and hydrogenated in an autoclave over a rhodium cata-lyst (5% Rh/C) at 30 bar and 40 C. After i2 hours, the product is filtered off from the catalyst and the filtrate is stirred with respectively 3 ml of cation exchanger (IR 120) and 3 ml of anion exchanger (IRA 410) for one hour. The product is removed from the exchanger by filtration and freeze-dried.

- ~ ..
' ~ :

: ~ `;
: ~

- 65 - ~0~42~2 Yi(-ld: ~.18 cJ ~92~ of theory) of a colorles~, ~moxpho~
pc)wder (cont~ining 6.7~ wat.~r per an~ly~is).

Analy.si3 ~corrected fox ~ter):
C 30.08 ~ ~ 27 N 8.~8 ~d 24.93 (Calcd,) C 30.01 H 5.34 N 8,78 Gd 24.86 (Foundl _ampl~ 10 Prep~ra~ion of ~ Solution of the Gadolinium(III) Complex of 3,6,~,15-Te~ra~a~icyolo~9.3,1]pentadeca-1~1$),11,13-t.riene-3,6,~tris[a-(2-hydroxyethyl)acetic Acld _.____________~____________________________.________~____ 1.0 ~a) 361 g (0. 5 mole) of the ~omplex obtained ~cording to Example l(b) (water content: 8.3%) is disso~ved in 500 ml of wa~er pro in jections (p.i.) with sli.ght heating. After adding 0.8 g of trometh~ine, ~hc ~olution is filled up wi~h wa~er p.i. to 1000 ml.
The solution is subjected to ultra~iltration and dis-pen~ed into bot~les. After heat-steriliz~tion, the solution is ~eady for use for c31agnostic purposes by paren~eral administra~ion.
~b1 The solution obtained ~y ~ltra~ltra tion a~cordi~lg to Example lO(a) is fill~d under sterile con~i~ione in~o multivials and lyophilized. Af~er ~c3di~1g th~ desired amount o water p.i., the admini~t~a-tion dose sui~able for int.rasti~ial inj~ction for r~iation ther~p~ i~ o~tained.
~'he proceding example~ ~an b~ r~p~atad With : oimilar success by substituting the generi~ally or specifically ~escribed r~a~tants and/or opera~inq conditions of this invention ~ox those u~ed in the precedlng example~.
From the foregoing descript$on, one &killed in the art can easily a~certain the e~sQn~ial characteri~tics of thi& inVention~ and without departing ~ro~ 'che :3piri~ and scope ~bereof~, oan raake various ahanges and modif$ca~ions of the invention to adapt i~ to varlon~ u~ne~ anA cc-n~t.~en~.

Claims (19)

1. A macrocyclic compound of Formula I

(I).

wherein a single or double bond;
Q is a N atom or NH;
X1 is H, -(CH2)n-R1 or ;

n is 1, 2, 3, 4, or 5;
m is 0, 1 or 2;
R1 is H or OH;
X2 is X1 or ;
k is 0, 1, 2, 3 or 4;
? and q n are each independently 0 or 1:
R2 is H, C1-C4-alkoxy, a functional group, or a bio- or macromolecule bound via a functional group;
A1, A2, B1, B2, C1, C2, D1, D2, E1, E2, F1 and F2 are each independently X2;
G is R2 or a second macrocycle, bound via K, of general formula II

(II), K is a direct bond, a bis(carbonyl-amino)group (-NH-CO-CO-NH-), or a C1-C4-alkylene group which optionally carries at the ends carbonyl (-CO-) groups, carbonylamino (-NH-CO-) groups or oxygen atoms, and which optionally contains one or several oxygen atoms(s), hydroxymethylene (-CHOH-), CH(X2)COOZ-substituted imino, acyl-substituted imino and/or hydroxy-acyl-substituted imino groups, and or one to two C-C-double and/or C-C-triple bonds; and Z is H and/or a metal ion equivalent of an element of atomic number 21-29, 31, 32, 37-39 42-44, 49 or 57-83;
wherein any or all remaining CO2H groups can optionally be present as an ester or amide; with the provisos that the 12 ring substituents A1 through F2 stand for at least 8 hydrogen atoms, X1 and X2 stand simultaneously for hydrogen only if at least one of the ring substituents A1 through F2 is not H, and that the macrocycle of general Formula I contains no more than one bio- or micromolecule; and physiologically acceptable salts thereof with inorganic and/or organic bases, amino acids or amino acid amides.

2. A macrocyclic compound of claim 1, wherein said compound is of the Formula III

(III), wherein is a single or double bond;
Q is a N atom or NH, X1 is H, -(CH2)n-R1 or :

n is 1, 2, 3, 4, or 5:
m is 0, 1 or 2;
R1 is H or OH;
X2 is X1 or ;
k is 0, 1, 2, 3 or 4;
? and q are each independently 0 or 1;
R2 is H, C1-C4-alkoxy, a functional group, or a bio- or macromolecule bound via a functional group;
A1, B1, C1, and D1, are each independently X2;
G is R2 or a second macrocycle, bound via K, of general Formula IV

(IV).
K is a direct bond, a bis(carbonyl-amino)group (-NH-CO-CO-NH-), or a C1-C14-alkylene group which optionally carries at the ends carbonyl (-CO-) groups, carbonylamino (-NH-CO-) groups or oxygen atoms, and which optionally contains one or several oxygen atoms(s), hydroxymethylene (-CHOH-), CH(X2)COOZ-substituted imino, acyl-substituted imino and/or hydroxy-acyl-substituted imino groups, and or one to two C-C-double and/or C-C-triple bonds; and is H and/or a metal ion equivalent of an element of atomic number 21-29, 31, 32, 37-39, 42-44, 49 or 57-83;
wherein any or all remaining CO2H groups can optionally be present as an ester or amide; with the provisos that X1 and X2 stand simultaneously for hydrogen only if at least one of the ring substituents A1 through D1 is not H, and physiologically acceptable salts thereof with inarganic and/or organic bases, amino acids or amino acid amides.

3. A compound according to claim 1, wherein Z
is H.

4. A compound according to claim 1, wherein at least 2 of the substituents Z are metal ion equivalents of at least one element of atomic numbers 21-29, 42, 44 or 57-83, or of at least one radionuclide of an element of atomic numbers 27, 29, 31, 32, 37-39, 43 49, 62, 64, 70 or 77.

5. A compound according to claim 1, wherein K
is or a direct bond.

6. A compound according to claim 1, wherein R2 is a functional group and said functional group is NCS, NO2, OH, NHNH2, NH2, NHCOCH2Br, NHCOCH2Cl, CO2H, or CON3.

7. A compound according to claim 1, wherein the bio- or macromolecule optionally contained in R2 is an antibody or antibody fragment.

8. A compound according to claim 1, wherein the bio- or macromolecule optionally contained in R2 is a protein.

9. A compound according to claim 8, wherein said protein is albumin, globulin or lectin.

10. A compound according to claim 1, wherein the bio- or macromolecule optionally contained in R2 is a polysaccharide .

11. A compound according to claim 10, wherein said polysaccharide is amylose, dextran or dextrin.

12. A compound according to claim 1, wherein X1 is CH20H, CH2CH2OH, or CHOHCH2OH .

13. A compound according to claim 2, wherein X2, A1, B1, C1 and/or D1 each independently are CH20H, CH2CH2OH, CH2OCH2C6H5, CHOHCH2OH, CH2C6H4OCH3, CH2C6H5, CH2C6H4O(CH2)3COOH, or CH2C6H4NCS.

14. A method of NMR diagnostics, X-ray diagnostics, radiodiagnostios , radioimmunotherapy or irradiation therapy comprising administering an agent wherein said agent contains at least one physiologically compatible compound of claim 4.

15. A process for the production of a macrocyclic compound of general Formula I

(I), wherein is a single or double bond;
Q is a N atom or NH, X1 is H, - (CH2)n-R1 or ;

n is 1, 2, 3, 4, or 5;
m is 0, 1 or 2;
R1 is H or OH;
X2 is X1 or -(CH2)n-(O)?-(CH2)k-(C6H4)q-R2;

k is 0, 1, 2, 3 or 4;
? and q are each independently O or 1;
R2 is H, C1-C4-alkoxy, a functional group, or a bio- or macromolecule bound via a functional group;
A1, A2, B1, B2, C1, C2, D1, D2, E1, E2, F1 and F2 are each independently X2;
G is R2 or a second macrocycle, bound via K, of general Formula II

(II).

K is a direct bond, a bis(carbonyl-amino) group (-NH-CO-CO-NH-), or a C1-C14-alkylene group which optionally carries at the ends carbonyl (-CO-) groups, carbonylamino (-NH-CO-) groups or oxygen atoms, and which optionally contains one or several oxygen atoms(s), hydroxymethylene (-CHOH-), CH(X2)COOZ-substituted imino, acyl-substituted imino and/or hydroxy-acyl-substituted imino groups, and or one to two C-C-double and/or C-C-triple bonds; and Z is H and/or a metal ion equivalent of an element of atomic number 21-29, 31, 32, 37-39, 42-44, 49 or 57-83;
wherein any or all remaining CO2H groups can optionally be present as an ester or amide; with the provisos that the 12 ring substituents A1 through F2 stand for at least 8 hydrogen atoms, X1 and X2 stand simultaneously for hydrogen only if at least one of the ring substituents A1 through F2 is not H, and that the macrocycle of general Formula I contains no more than one bio- or micromolecule; and physiologically acceptable salts thereof with inorganic and/or organic bases, amino acids or amino acid amides, said process comprising splitting off the blocking groups conventionally from a compound of general Formula I' (I').

wherein G1, X1' and X2' in each case stand for G, X1 and X2, the hydroxy groups and functional groups contained therein being present in the blocked form and, respectively, as a precursor, and Z' means a hydrogen atom or an acid blocking group;
optionally generating the desired functional group;
optionally reacting the thus-obtained complexing compounds of general Formula I wherein Z means hydrogen in a manner known per se with at least one metal oxide or metal salt of an element of atomic numbers 21-29, 31, 32, 37-39, 42-44, 49 or 57-83; optlonally bind1ng the functional groups to a bio- or macromolecule; and subsequent optionally substituting any still present acidic hydrogen atoms with oations of inorganic and/or organic bases, amino acids or amino acid amides; and/or converting corresponding acid groups entirely or partially into esters or amides;
wherein the complexing can take place before or after the splitting off of the blocking groups for the hydroxy groups and functional groups and/or generation of the functional groups and linkage to a macro- or biomolecule.

16. A pharmaceutical agent comprising at least one compound according to claim 1 and a pharmaceutically acceptable carrier.

17. An agent according to claim 16, further comprising at least one additive customary in galenic pharmacy.

18. A pharmaceutical agent according to claim 16, wherein said at least one compound according to claim 1 is in the form of liposomes.

19. A process for the production of a pharmaceutical agent according to claim 16, comprising dissolving or suspending the complex compound in water or a physiological saline solution, to bring it into a form suitable for enteral or parental administration, said agent optionally containing additives customary in galenic pharmacy.