FR2617843A1 - ANTINEOPLASTIC SUBSTANCE DITE ROLLINIASTATIN 1 - Google Patents

Abstract

A new substance, having a potent action of inhibiting cell development, has been obtained from a South American tree, Rollinia mucosa. This substance has been named rolliniastatin 1. Physical and chemical parameters are described, including its ability to strongly inhibit the development of N.C.I.P388 lymphocytic leukemia. Pharmaceutical preparations are described containing rolliniastatin 1, its synthetic counterpart and their pharmaceutically active and non-toxic derivatives, as an essential active ingredient of these preparations.

Description

The present invention generally relates to a new substance

  antineopiasis hereinafter referred to as "rolliniastatin 1", its synthetic counterpart, pharmaceutically acceptable derivatives of this substance and the

  treatment of a host with the disease by them.

  This research was partially funded by CA-30311OlAS grant from the National Cancer Insti- tute, DHHS. The family of <200 species of advertisacee) is composed of shrubs and trees mainly found in the tropics. The genus Roil {nia is generally found in Central and South America and R. mucosa (Jacq.> Vaill, also known as Annona mucosa (Jacq.), Was used for medical treatment of tumors by natives in the West Indies and In Indonesia, in order to evaluate the possibilities of use of this plant in terms of potentially effective antineoplastic components,

  Rollinia mucosa were collected in Guinea in 1979.

  2C The lipophilic extract of these seeds showed activity in the system of P388 lymphoid leukemia and the fractionation of this extract was guided by tests

  using the murine P288 system in vitro.

  The present invention relates to the discovery of a novel and effective chemotherapeutic agent which is extracted from Roilinia muco-a seeds in the manner described hereinafter in detail and which can then be formulated into effective pharmaceutical preparations prior to demonstrable and confirmed levels of efficacy. . ' anti-cancer activity, when measured by generally accepted protocols, in use at the United States National Cancer Institute. The main substance of which

  it is here question was referred to as "rolliniastatin 1".

  The present invention further relates to the preparation of a synthetic pendant of rolliniastatin 1 and pharmacologically active and nontoxic derivatives thereof. The main active ingredient of the present invention, namely, rolliniastatin 1 has the OH CH structure, and has shown surprisingly potent activity in inhibiting the development of the

lymphoid leukemia NCI P388.

  Accordingly, it is the main object of the present invention to provide a novel agent useful in slowing the development or remission of one or more

of several types of cancer.

  Another object of the present invention is to provide methods and methods for isolating an antineoplastic substance from flora in a form in which it can be used easily and usefully in the treatment and therapeutic care of one or more types of cancer. that occur in human haste. The present invention further aims to provide means and methods for making pharmaceutical preparations useful for the treatment and care of neoplastic conditions, preparations which contain, as an essential active ingredient, a factor extracted from Rollinia seeds. mucosa, its synthetic counterpart or a pharmacologically derived

active and non-toxic.

  These and other objects which will become apparent hereinafter are readily achieved by the present invention in a remarkably unexpected manner, as will be appreciated

  account easily from the detailed description that

  follows from an embodiment chosen for

example.

  The depletion of Rollinia mucosa seeds (600 g) with hexane gave 136 g of an oily extract, from which rolliniastatin 1 was separated by a series of chromatography operations. Size exclusion chromatography was followed by gradient elution chromatography on silica gel with hexane-ethyl acetate. Rolliniastetine 1 (1.75 g, 0.29% yield) was isolated as a wax, m.p. 81-83 ° C from acetone; [a] +25,2, (a] s7e +26,2 ', [cse +30,1', [aà] s +48,5 ', [] (XI +76,70 (t = 28 C , 1.03, CH 2 Cl 2); UV X 1 (CH 3 OH) 224 nm (log λ 3,93) (α, β-unsaturated carbonyl); - (CHCl 3)

  3580, 3460 (OH), 2928, 2855 (C-H) and 1748 (γ-lactone a, 3-

  unsaturated) cm- '; SP-SINS, m / z 645 [M + Na] +, 730 [M + Ag]; and

  HRMS, m / z 623.4917 (calculated for C37HesO7 + H, 623.4887).

  In deuterioglycerol, SP-SIMS gave m / z 627 [M + D] +,

  which indicates three exchangeable protons.

  The spectral features indicate that rolliniastatin 1 is a member of a new class of bioactive bis-tetrahydrofurans ("acetogenins") that includes uvaricin, rollicinin, isorollicin, desacetyluvaracin, chérimoline, dihydrocher-iminol and asimicine. These compounds typically contain 37 carbon atoms and include a (generally unsaturated) lactone residue and two tetrahydrofuran rings to which a long

hydrocarbon chain.

  The 3C and 3H NMR spectra presented in Tables I and II below, elaborated by COZY and heteronuclear correlation experiments, established the existence of the γ-lactone α, ε-unsaturated residue of rolliniastatin 1. The following characteristics were supported by the lC offsets: the C-1 carbonyl group (6174.5, singlet), the double bond between C-2 (6 131.1, singlet) and C-35 (6151). , 7, doublet), a doublet 78.0 for C-36 and a 19.1 quartet for the C-37 methyl group. A heteronuclear correlation experiment attached C-35 to a proton field

  descending to 6 7.17 and C-36 to another proton at 6 5.06.

  Chemical shifts suggestive of fatty acids have been observed: 'H and' C signals (Tables I and II) for C-32, C-33 and C-34 and for a number of

  approximately equivalent methylene groups.

TABLE I

  CHEMICAL OFFSETS OF RM; H AND COUFLAGES OF FPROI'CS

FOR ROLLINIASTATIN 1

  ## EQU1 ## where: (J (Hz)) 3a 2.50 dddd 3a, 3b 15.1 21 3a, 4 3.5 22 3a , 3, 85 3a, 35 1.5 3b 2, 36 dddd 3a, 3b 15.1 3b, 35 1.5 3b, 4 8.1 26- 33 3a, 36 1, 6 3b, 35 1.2 34 3b, 36 1.4 3b, 36 1 4 35 7> 16 ddd 35.36 1.5 4 3j85 dd 3a, 4 3, 5 36 5/02 dddq 36.37 1.5 3b, 4 8.1 37 1 41 q 1455 m 6th-13 1, 25 14 1.50 m 3, 38 m 16 3 85 m 18 1.7-1) 9 m

19 3 38

3.85

  The chemical offsets in ppm (6) (at 300 MHz) are given relative to t-tramethylsilane, with use

  of a solution in deuterochloroform.

TABLE III

CHEMICAL SHIFTS OF NMR-C DE

THE ROLLINIASTATIN 1

  6c & g Offset of Chemical Chemical Chemical Cleavage 9 # C (a) oc (6) # C (e) 1,174.5 s 15 74,0td 25 32t8 + t 2 131,1 s 16 83t0 d 26 25,5 t 3 33.2 t 17 28.7 t 27 4 69.9 d 18 27.8 t 31 6t 37, 4 t 19 81.1 d 32 31t9 t 6 26.0 t 20) 81.0 d 33 22t6 t 7 29.5 21 27.8 t 34 14tl q 12 29.3 22 28t4 t 35 151.7 d 13 25t7 t 23 83, 0 d 36 7-79 d 14 34.1 + t 24 7118td 37 19, lq

  t These offsets are interchangeable.

  The spectra were established in solution in deuterochloroform and the chemical shifts are

  expressed in ppm (6) relative to tetramethylsilane.

  An NMR DEPT experiment revealed eight '3C doublets associated with oxygenated carbon atoms (6

  69.9, 71.8, 74.1, 77.9 and four others 81.0-8: 3.0).

  One of them is in the lactone nucleus and the other three have hydroxy groups that explain the three exchangeable protons. The presence of three secondary hydroxyl groups was confirmed by observation

  spectral changes caused by acetylation.

  The remaining four oxygenated carbon atoms were attributed to the tetrahydrofuran rings. Methine proton resonance, supplementing each of the doublet C shifts, occurred in the region 3.38-3.85,

  except for the C37 proton signal at 6 5.02.

  A low-resolution mass spectrum of rolliniastatin 1 gave a strikingly parallel pattern of fragmentation to that of rollinone, bis-tetrahydrofuran isomer, which has a 14oxo group and a saturated lactone residue. In addition, the fragmentation model of the mass spectrum made it possible to locate the presumed bis-tetrahydrofuran system with its two α-hydroxyl groups between C-15 and C-24. A

  evidence confirming the tetrahydrofuran system

  hydroxylated was provided by 1H NMR. COZY experiments have established couplings between the following sets of protons: H-14-H-15 (6 3.38-1.38), H-15-H-16. (3.38 - 3.80) and H-19 - H (6.38 - 3.88). Relationships between H-23, H-24 and H-25 were discerned by the same method. The following C-13 / H-1 connections were plotted by heteronuclear connectivity experiments: C-14 - H14a + 14b '<* 6 34.1 -i 1.44 +

  1.50), C-15 - H-15 (S6 74.0 - 3.42), C-16 - H-16 (633.1).

  3.86) and C-19 - H-19 (6 81.0 - 3.88). Corresponding connections have been noted among protons and

carbons from C-20 to C-25.

  The plausible hypothesis that rolliniastatin 1, as well as rollinone, has an oxygen function at C-14 has proved false, despite their similar mass spectra. However, by means of NMR experiments (including COZY, heteronuclear correlation and PANIC simulation), the third hydroxyl was localized with respect to the lactone ring and located at C-4. Allylic couplings (I = 1.5, 1.2 Hz) between the two non-equivalent protons 3a and 3b <6, 2.50 and 2.36) and H-35, 165 confirmed this attribution. In addition, the protons: 3a and 3b had long-range couplings (i = 1 ,, H'I'N np p TpueGs saloDo4ood sap uoTvslTIT; n oaAq 'lm / gd _-OI x S'b = 5Hq! 99u , P% 9E ap% t% ugauoJo-d of a-1eledde qTe where iS suT:% - sQTuTlliO vl ap anbT2oioTq uoTlenIvapq agaXuas9zd I auT; ezsTuTIIoa eI ap SXIH, P uoTîequaSemi; -el; d 'pa9 4ao in the south of the world, it is necessary to have 61% of the population of the country and of the country and to have a high level of education, and to be able to meet the needs of the people of the world. Tnb xnso e sanSzuapT wuaajlianqlTA Iuos 61-t5 sauSvvD sal -nod auToTuTsel ap O ap seovIvop Sa u'Sl-3 uaos' - u 4Tos' elz9; TTp UoTIn2TiuocOE el% uop sezaosIoasp sap% nuos sisodmoo xnap salt an5: uanbTpuT suoTieAisqo sao ' (SZ-Oa QL - 'tT-3 -uesnioedsal) EO zD' BL-, tusulsouoo Tn5 xnao ap uoTdaoxlI IsaieTlTmTs nad anblanb% uos) uToFiTsvl ja (II nealqe-%) I auteisPTuTIlo -el -Inod 0, ap? sa2l2Po; Ppp saq [(g96T) ú611 'Ve' salofooiaeH 'o] slnaejoqeljoo sas% e% qoeaddnd ied 4T-oap% suaUm-ol 'auToTUsT5 aafauosl auenlnozpqDar-mIq aI nod elquieg - + uaslae53 -sa aTuTI; p ua or aqooa% aun a) Ae 1 auT: 2-s2TuTIIoa eI znod sa1dv- T aueszdel aznonlas -e Ol <S '- Sla g) S-3 ap 14a (V'úú - TS'Z + 8 &' Z 9) ú-3 ap neaATu suo% ozd ea suoqoec F% ua sa 'Tdodde suoTxauuoo sap Tiqe a 4QQ IIQQ IIuQ2.ixo a -9 iax ;; 4TQ, 2b, - anb aaqhu, l X.a unl - uatnbpui H '. % 4 B ap ap Sa2i1oDEp salt 'TsUTV' 6'69 9 I xalqnop a asuoo 4uessTeiedd2 auoqoeD e aoauuoo -V-Ha 'inol ucs EC, nb; @ke; suoc) Axis II (8 & 9) b -H e (ZH T "9 'b', = -) saIdnoo 4uaTeta qu 4a ee suolozd sae anb e-eiuoo. SsnQ e II '(cO' 9) 98-H oen (ZH b'l ú8 8 9Zz 171 z 141 - 311 (100) CH 3 -22 21 17 CHoCHH 2) 11.

OHI '1OH

  The biologically active Sluc oside derivatives of rolliniastatin 1 are obtained by copulation of the principle

  with properly protected sugars or else--

  hydroxyl compounds, by applying methods known

  in the art [see Rethods, in Chemistry, R.L.

  Whistler and J.N. Bemiller (editors),

  Academic Press, N. Y., 1978, Vol. 6, to 'The Carbohydrates-

  Chemistry and Biochemstry, W. Pigman, Academic Press, N.Y., 19811. These derivatives of the rollinastatinq 1 are

  used for the same purposes as roll iniastatin 1.

  As has been demonstrated, rolliniastatin 1 has free hydroxyl groups available for derivatization. Thus, acyl esters of these compounds can also be prepared by methods well known to those skilled in the art. Acyl derivatives of rolliniastatin 1 may be

* I I OH,

I! i I

I I

  Used glucosidiological derivatives that are activeomposs of the departure. As acids which can be used in the acylatin of a rolliniastatin 1, are obtained by copulation of the principle with suitably protected sugars or other hydroxyl compounds, by applying known methods.

  in the art [see Methodes, in Chemistry, R.L.

  Whistler and JN. Bemitler (eds), Academic Press, N.Y., 1978, "Vol.O, oul he Carbohydrates: Chemistry and Biochemstry, W. Pigman, Academic Press, N.Y., 1981] 3, These derivatives of rolliniastatin 1 are

  used for the same purposes as rolliniastatin !.

  As it has been shown, roliiniastatin 1 has free hydroxyl groups, available for. the formation of derivatives. It is thus that acyl esters of these compounds can also be prepared by methods well known to those skilled in the art. Acylenated derivatives of the iniastatin 1 roll may be used for the same biological purposes as the starting compounds. Acids that can be used in the acylation of a rolliniastatin 1 include: Kxoq %% maTdmsxe auoqjo ap samo4Q xTs ap snld si2d: + ur ... oduioo to inaT-ii} uu xoz) the : wuaumeqou 1-inaT-ip; uT A :: ooI1 no ouxc'OoT q 'oueoA'ouTm' olo 'XXOlpKq o-iTu suacadnol5% pq sauaBoeq, p samo-e sap sanaTsnld no one led san% ;; sqns: uos Tn5 snssap-To saeTo spuoqlmoolpXq sanbTIxoqieo sapTDo sal 'ssinTsqns-Kxooiv% a -ouAdTy% -Ou-QD -_ouTua' -O;, 4P '-, xXOlpxq' -14T -OUEIROIuq seuoqieoo.apKq sanbTiXxoqaQo sapToe a0moo ': aueuuaTAuoo -saTIlTuiTs no snS;%; oDeK ^ deu spT; e, the anbSIodo-dikuaqd ap e, the anbTureuutT'D apTo, the anbTaIPaTIuaqd apTo, the anSiuoTdouRdd uRdd epTzeI a eanbST: paUpqd apT0o, l aldwexa -1d 'ssnSnTqdTI -sanST4 = oiQ sanSTIKxoqivo SepTOQ sap (a)% s' sasi-elTs no enb! ozuaqlXnql qIa p5oe, the anblozusql KNQOSITT BPTPIoQ 'ilozu qlKqa 6ptoei anblo4qdeu apTDo, i sanbTnIo% apTo, the anbLozuaq apToe, the aidmaxa ad' sanb ; I% = oae sanbtIXxoqmeD sapTov sap (p): sa-iTelTTs no snbtaseuexaqoiooifqa a PIo, I anbtlKnqsuexsqxoqoIoXo apToeI 'anSI% oaseuexaqoiDoo PFo, I' anSuoTdoldaueusdoIoXoD epTQl 'snb5: ge0 -aue-uadolOXo' sapIe, I aldusxa ed Isp-ingesuT no snes sanbI; eydII-ssnbTIOOIe sanbTIXxoqiuo sapToe sap (o): sa-ITFsITS no anbTKlxoqeoauexaqoloKoKlXdodTp BpToVi anSIxoqJeoauexsqol DúO lKqmlp spToe, I. 'anSIKxoqlo -auexaqosIOKO apToe, the IKxoqleoauap Z uadoIoKoInxOqll op3 T,' in -Ixoqloeapu9% uedoIoO pToeI 'sanbTTlxoqaQo -aueXuadoloXO apTQi sanbîiXxoqvosauuqnqoloXo apIo, e. aldmuaxe of ssansuT no sa-n.es senbTlooTúie sanbIxoqieo sEapIo sap (q) sslIms no anbiouXzo anblouK4daq 'an5! ouxxaq' anbTE9o 'anbTualIXopun anbiuokoao' anbTJolanSIQ, 4s' sanbSigoSm nbi i -ed 'snbouQoripeuad' anbS; sTiXm lanb5ouvoapT% lanbTlnvl 'anbIouQo,) pop' anbSiouuoiFp 'anbTIldwo DnbSodeo' anbTui2IAOSl 'anbST, a' anbT4ipoQlx-n '- its% anbjl - zKnqosI' anbiKqnq 'anbTuoidoid anSbT4iaoQ sapToe úal aldmexe -1ed zaa; mlno a, 4To-p auleyD 1? 's - i, 4.2fuNoNoS sHaBlEr sHaBlEr sHaBlErS

ú8Z 9Z

  g2 'auTuTwe, eUlolTç gl'btbAnd apToel anbTuTTnAgT ANbAt% oost aploeI' anbTaTo aptoe, the anbTIem apIoe, I (aqrp aneuoqicoouapoapxq) enbTmiojXxoqnp epF, a1 anbizoel aP! O, the anbTuo! doudoueio apTov, the InbT $ ooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooo -ud- Ja - the f'_oO 'ao.1 Sanb 1 1TZUEqojo tIo p-. * t 'to apToe, the ebmUTXxt ^ = apoil lanbozusqúxoqpmIa4 aplTogI anbia: Fi2 apI: lz, snbTII9 apToil' anbTIXoaosga -q apT: It, enb1ozuaqKxoapxq-d aptle 'anbTIfOIT S 5Z apT,' Qa 'anbIsTue apToDI' anbEIozusqo o qo -d% a -ur -o apFO3, the anbTsTIua2omoq epTou, lEbElxoqLoau-exeq _oio] KolXqaurT2% _g'g-o ql, maPToe, 1 lanbITIKxcqcioquvqued _OloAolAN; dmTl ± ú'ZiZ- = lq- & apTousi lanbTIA 'xcqa-ec _auexaqolOlOittX'qs1ll -; _ ooqip-E'I apTIo, l'anbTlXxoqieo Oz _ouexaqoioKotKql-Dva = - ooqIp-g'I aPtO, t' anbtIxoqapo auexaqoimioi /% a-gouroq-g sPTO, * l'anBtIKxoqeo _auexsqoioAoiúqX = _ú_ = o2q-ú sptoeI 'menbTIxxoqavo -auexeqo lOfOiKq% -_ 0_ TP-9 Ir-'5 pTo.1' menbIIxoqavo _auexuoioXol- \ A: DlKq lgo2qTp-5 'b apToQ.I' enbTlXxoqa 5 _uxsqolOXOIiqeg-ooq I? ## EQU1 ## where ## EQU1 ## 1 enbTIA xoqivo _auexaRoloKOI @ Rqam-g-OoixqIp-g'g apTo, .I 'anbTIi / xoq12o _oupxwqolOmlASqua = -go = oq-9% a -g saptoou.1 anbTIxoqu o _uuxmuolOXOIq% = - g-_o = loq-g % a - aqtxxqqqqqqqxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx + Eu epTla.1 'anbTm-irIqs ap eI e nS IIxmqiesauexsq I f o0 Iq - _z apTo-, I anbTU0112e ^, u apToQl ienbTipl eAopoT-g-q _) D aPToesi anbTaX-nqouioaqX -) O API-e , AnbTuoTdoidoolqo5 - '% a -. epToeI 'anbI% 43ei0olqoIF% -TP' -oror apTme, .I :; edteaxa id% uos 5, nTsqns 'UOqlOOap sanbt lAxoxcqi3sapIZe its aa -its-lQosT sam0o: SnaI% -' A>: oIixaq 'KXOiQ' KXO% nq 'fxodoid' fxoq% L9

2 6 1 7 8 4 3

  valine, isoleucine, leucine, phenylalanine, proline, serine, threonine, tyrosine, hydroxyproline, ornithine, lysine, arginine, histidine, hydroxylysine, phenylglycine, paminobenzoic acid, m-aminobenzoic acid, anthranilic acid, aspartic acid, glutamic acid, aminoadipic acid, glutamine, asparagine or the like. The administration of rolliniastatin 1 and its pharmacologically active and physiologically compatible derivatives is effective for the treatment of animals and humans with a neoplastic condition, e.g., acute myeloid leukemia. acute lymphoid leukemia, malignant melanoma, lung adenocarcinoma, neuroblastoma, small cell lung cancer, breast cancer, colon cancer, ovarian cancer,

  bladder cancer or the like.

  The dose administered will depend on the nature of the neoplastic condition, the type of host affected, including its age, state of health and weight, the type of simultaneous treatment, if any, of

  the frequency of treatment and the therapeutic ratio.

  By way of illustration, the dosage levels of the active ingredients administered are as follows: intravenously, 0.1 to about 20 mg / kg; intramuscularly, 1 to about 50 mg / kg; orally, 5 to about 100 mg / kg; by nasal instillation, at about 5 mg / kg; and by aerosol, 5 to about 100 mg / kg of weight

body of the host.

  In terms of concentration, an active ingredient may be present in the compositions of the present invention for topical dermal, intranasal, pharyngolaryngeal, bronchial, intravaginal, rectal or ocular use, in a concentration of about 0.01 to 50% by weight. / p of the composition, preferably about 1 to 20%, w / w of the composition; and in the compositions for use

2 6 1 7 8 4 3

  parenteral, in a concentration of approximately 0.05 to 50

  p / v of the composition, preferably from about

  w / v. The compositions of the present invention are preferably presented, for administration to humans and animals, in dosage unit forms, for example tablets, capsules, pills,

  powders, granules, suppositories, solution = or-

  sterile injectable suspensions, non-injectable solutions or suspensions, oral or similar solutions or suspensions, containing appropriate amounts

of an active ingredient.

  For oral administration, solid or liquid dosage unit forms may be prepared; The powders are prepared very simply by spraying the active ingredient Up. a sufficiently fine particle size and by mixing with a similarly sprayed diluent. The diluent can be an edible carbohydrate such as lactose or starch. A sweetening agent or a sugar is

  advantageously present, as well as a flavoring oil.

  Capsules are produced by preparing a powder mixture as described above and filling molded gelatin cases with this mixture. It is advantageous to add to the powder mixture, prior to the filling operation, a lubricant such as talc, magnesium stearate, calcium stearate or the like, serving as an adjunct for the filling operation. Soft gelatin capsules are prepared by machine encapsulation of a paste of active ingredients with an acceptable vegetable oil, a liquid petrolatum

  light, another inert oil or triglyceride.

  Tablets are made by preparing a mixture of powders, granulating or pelletizing, adding a lubricant and pressing into tablets. The el anb Té is prepared. uoTsuedsns ap + uai2e a, p apTel e 'aTa-do-1d of aTno; qiA a salaLosuT.suLio oaAe; sep ped 5aIsdezd aex% uaAnad suoTsuadsns sea' uusTmoi aue2Q a his sETidoiddQ su-eoolnpp sep oate anbTIoote-oapKq elntqEnA 4uwsIIlln ue a2dgd 4sse iTx $ i a dolTs an emo-c; Inod'uoTiaesuoo ap suuea2sep e usao sua3Q sep 'eaionsnp oaA xnanbe alnoTqqa a suep Oç

  sariossTp eal.:uaAnad nea, supp salqnlos seaao se-

  uaT-Tsoduioo uuuooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooh ap saiedlzd ea aAneed suoTsuedsns -ke jTxa 'sdoTs anb salla alGao U0 Ti% STUTiPI Z inod saepnbTI sealT; Tun sanbTSiolosod saiao; seaj eqneu. oo ep ailTo ue iodod apddolaAua aun, pa asoinlIeo.I & qm ape eaeyspddolaAue eun, p ennbul - amomIue aeuulsakeuT uoTIdIosqe anod c4uemae; aal a no qou, p oz 4ueUmaegalun ied arn; T; suoo aoTjjoaold addolaAue eun, pTue2% ueaasne24uAe ; . 4ed pawTdmoo aq simTjdmoo us, ssald a; Tnsue -sa, T $ qni Buelui elq a'W-EuTm alTnq aunp no oiye ep 'snbIipas ISs ap' anbI; s apT-o, p uoT; Ippe 5 edu aeulTdutoo sap uo; eImoj ap selnom xnu aoueaiqpv

  neal a-ia ,, p UT; e ssT; TjTqnI ea; quaAnad saeIIT;

  (saeIITSed) sueus29e; ua ssTaq as-i; s uaaned; ue "inspl Sgmlo; ues%; eItdmI srTidmoo salt asnaTit% e; sed el saAe- - assed AE4 4ned IT AETP nb-V-; sa o 'a91T; South has 01% 4p 4nad apodnod ap a2ujim to the uoTelnue-S -e ap eauv.-n ap aa; 4 V alqTao a sJa-a%. W asogo ap jassid uQsTQT el us 4a ouTo, p seluTonm a no solnlalq Ep uo; nTors aun uaTiTiTiTiTiTiTiTiTiTiNiNg iNTeNTiTiNgS iNTiNiTiNiNg a: oNiNiNg oF tHe oiTiNiNg iNTiNiTiNg iNTeNTiNTiTiTiNG iNTeNTiNTiTiNG oNTiNTiNTiNG iNToNTiNTiNG iNToNTiTiNg Aseq 'a no uanlTp a oETA' EsTAiAlnd uauialquaAuoo I fuaiTpauT a 4uQeau2jim ua saupnodap a2ulpim úy8L 9Z

26 1 7 8 4 3

  gum arabic, tragacanth, methylcellulose or the like.

  For parenteral administration, forms

  Liquid unit doses are prepared using an active ingredient and a sterile vehicle, water being preferred. Depending on the form and concentration chosen, the active ingredient can be suspended or dissolved in the vehicle. To prepare solutions, the water-soluble active ingredient can be dissolved in water for injection and this solution can be sterilized by filtration before filling a suitable ampoule or vial and sealing it. Adjuvants such as local anesthetic, preservatives and buffering agents may be dissolved in the vehicle. Parenteral suspensions are prepared in substantially the same manner, except that the active ingredient is suspended in the vehicle instead of being dissolved, and sterilization can not be effected by filtration. The active ingredient can be sterilized by exposure to ethylene oxide before being suspended in the sterile vehicle. A surfactant or wetting agent is advantageously incorporated into the composition to facilitate uniform distribution of the active ingredient. In addition to oral and parenteral administration, the rectal and vaginal routes may be used. An active ingredient can be administered by means of a suppository. A vehicle that has a melting point around body temperature or a vehicle that is easily soluble can be used. For example, cocoa butter and various polyethylene glycols

  - (Carbowaxes) can serve as a vehicle.

  For intranasal instillation, a liquid unit dosage form is prepared using an amino acid, or the like, and the salt thereof is treated with a salt thereof. samlialTno 5u saI 'smqo sal' lapnod ap s: + anbed sal 'sa-to4Tsoddns saI' seliTTsmed salt 'salnsd-e sal SsmTmoo Sal -.uos uo! + ueauT aeuaspad Il uoles samiadoaddo e saecu: Tn sanbT2olosod semiot ap saldmaxe sae' uoTIueAuT suesa-d m ap% SmnasTaoo sap -uanTsuo + Where has seaQIa o sao ap saluipuadpp euemapoaaTp OUCS sallallaahu - + a + IuodTiosEp has uaspa of el-suep aiesodxa, nb went bmoqI zaqo anbTnadvaEqq. a2esn e am4ou aouensqns elie% ^ aun + uvuauoo uoTITsodmoo AUNP Uote-eda-d el ap: _e, I suep aeuajpquT uot; vTuTil ei (q) 4a '$ @ iu .Sqo S -g _iajjnoT + led ENBT, 4nade2laql. 4AJ; a, l - a + aA-io-e-aug Qsqns -e ap sadojd SNSI-ST9, oelo ssl (1): led sa0oTp + uos UOT-uaAut auaspaid el ap Saat-e4Tun sanbi2olosod sanio; Salt + UeUja7uoo suoTeoTtToads ssq '5 s TlbbiTnaonumaqd alnoTqaA no uaTdT xa 0O

  % uunlTp al oaAe UOT, + eTOosse ua piaTspp anbTInadQjpqq.

  s + a; Fal elanpoid lnod asaInoIeo 'aIT4oe sou-esqns ap aouTma-zaisp0d ia; T: uvnb aun 4uuua: ouoo p, 4 $ anbeqo xnw = tue ^ suTeunq sapns sap -nod sanbTiolosod his $ Tunnel uuuaToUuTo inb sagai2das: uama @ IaTjpjim sp4i a C sap au2Tsip suoTioTpuaAa_ sal 4a uoTdTaosap eI suep aasTI $ + n , aTeTun enbT2olosod am-ot, uoTssa-dxa @, 'saeqeTsap no sal-esa a-+ a: kus ^ ned Tn5' s4uellTnom s-4uase% as% + ueAlos -oo anb IsI 'Slensn s- + ueAnpesaoaAe OLTRIES no auedoad a4ozul 'enbTuoqUeo apTapXquvI aueq4azUoIonltTpolqoqtp al aldeaxa ed' pTnSbTI no xneze. anasindold an oo uoTlsa-d snos losogae aquoq aune suep snqo e- + uagnad s; Toa s4uaTepguI eel ssloso9, ae p auio; uoT4STIIN in anod JsToqo- UoTq; STuTUp1, p ap s ssa uoIjiQ ;; nsu, anbsiol aeInuaot ea + 4nad aqoas a-pnod auh saua3old soaDue2sqns ap a + dmaxa nea in aouagataeld ap a, _Idolddp anbInoa.Ieqd Ianoqia a 4a; T7o2 + uaTpa39uj ú98Zi 9Z 0, ea Sg 'g ap sasop sap -ui2uaouo-us sandu sap s lqp qms uo.g; ap aedjid uo IaanppcDojd auIaux ie 4uLnbTIdd- uS xno ad sTo; aB; nb 5 has a sandsdvo :: nap no aun, p uo> 4STsTutwp2 nd enbTs-sdou uoloao; Eun, p -uaniaq-p; lnod satoo; ea uos saBnsdço sSa ^

  BI! Nlqiq BIs-uem -e ap glnsd-oua sTnd -

  uemuiT $ .uT i2uT-eIm qsa 4nol B 'BauT aapnod ua s4Tfnpi o0 s uTpBI2uT saelne xna -, no Q 4sa' IF F anasiuoeiotu a, p uaXoi n2 pSTATp quemaiuTi 'jT4o lueTppgauT, q 3 z mnTspus-em ap awsZ 9 z S12U ap uopTUV 2 OZ i5sTUOIOTOE fTq L quaTppLagul: s4uaTpa2uTp s4uiATs saT; uQnb: a Sad14. sap aTqaud s saa-Edvi d + uos 'fa; ua-Tppi2uT a, p 2SM oz - + us2U4uoo alnsdvo enb-qo' .aloDnq aboa -da 2sn F samTirad xnap ue AINP AUB + eui {ap salnsdo 0001t alnp auTslu2 ap salnsdgo Oz -: u uT: Tsoduroo senfxoq uou wa sJT: Of 4uuianbi4nfol uu1eqd siapT-ip sanal a fnlb:; q; us uepBuad uos' I BuTeseTUTTIol -IT UZTs2ep., T% UBTPi2UT.,

  uot4e4or l slanbsalI suep '.uBATns nb SBldeaxa sBl suep u 1.

  úB + TcIp 4uos salis 'UOTIUeAUT aBuespid el JasTICouoo inod saBIdaid uuo sanbt-oiosod saBUot slnaTsnrid alzimaxg To-al --Eao quania-l nu 4u4TU, 1 au B a uoTBuaauaT 9uBs;' id and ap saTie; Tun 0O senbTi2olosodsamLTo sap uo6oaTUoo Bal 4ua nilTnT suoTe suoTe -idald sB- 'sTlqe-, uatq sapaoo-d sp aed

  sBsIdad aBi; u @ Anand qe anbTuqcaOB ul suep salqIuodsTp -

  sBuI-sBIalla Iuos Inb sanb; nscvemaqid s-aoue4sqs sp uoT4esTIT4n oaAQ saBThTun sanSTfoiosod saBBLoi setllai ap snos simedaid% ueml'ToBe; a44uaAnad senbTsvldopuTs s + ua $ e aBurIai SaiOldma s $ sTaUtpaj2uT saB aTiom9m quesUs5d al suep saTaoap sUIo o; n; p n-irp Bb TsuToe &saIo; o ap anbuoolTnb

ú '8 L1 9 Z

  nor aed sTo; asewn aun smp; adioo xnap 5 no one, p algao uo: IeTu-mpeI id anb-s-edoau uoT ^ poaIo aunp ausas; e-sod: o @ uos uss smTdoo bag To usTpazSuT, T ap m Oz duueuuaouo iuTaduoo anbeqo 'sgmTadmoo ua spsseid sdnsua: uos sduPqInspa salnuv2 sa' 'I1 oU IlqTF OS a smaae; i? aojo ap a $ essed avd sapsTsq duos salITqsud sea1 alTT; s-ed 4Fe juamam; ITU; ga2uQjiPU 4SA sound al Stnd SmuaspemuI sa: no twelfth g: noPw: its AIT nassuoxoTm F a, p uaúoui ne- asTAip luauaauTI IJT: ^ i: uaFpip-igu-c, 9 q; l599I spInbII n% Io <^% a SZ S buinTsau2im ap asz; S! 2 UHFWLRQWHUHHQWHGHUHUHUHUHUHGHUHHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGHGH sep t%; ed F saedpad quos 'toe uaTpuipiuT Unp 5I OZ unqao 4uQuausuoo' sau $ dmoo 0001 O, uoTIsoduoD nop aed sToj s $ oi; F aun salnsduo xnap 5 no aun, p aeIio uotdkastuîupu, the aled anbTsQldopu uoTIosaTT aun, p ausmstd el -nod ssooITe; duos selnsduo saO snssap-To ainblpuT as / $ Tun el ap 4uulnsd-oua, l ua smnd salq-lnsdeoua aouuqsqns assao a-pua- -nod 09 asqiosmKod ap OO suep uoTsuedsns O0 a asodmoo sal s-% m ad 4uSuemmoo ua sasgdadgd uos (lFe -i nasmuolo-I unp uiXoi ne rosTATp lueuauTF) j $ i; o 4aapalxut unp 2u OZ 4ueuasuoo ai-eoonq ason lrd asmsn W asoliad alnas aun, p allom auT; ieiS s selnsdeo sme alloW auT, 4Ia, ap sslnsduo OE, UoIqIsodmoo unp 2 Og oc, gz 'g aed snssap-To alnaaot el suep OsEs% Tn 2 Oz sel% upSeladme ua FT:% o% kuaTpDgiuTp z% 8 [ú2 8gL19Z s; -uWFnTSsi4Tzunb; a sad sap UT1id aPdPad 4sa 'btseldoiu uoT-oa; fi aun, p% u Tw lnod TI4o 4uaITpra2uT one, 2m p 0 li GAPD quuatuuoo 'letQia-ue-sd uoT4BaPuT anod aTTJIZ4s esnenbQ uotTsuadsns aua aIqwr% has uT -4tnP LAR O aq; or: q.npo.xcj OC

"-g uoT4T-sDd1Ioc-

  ino: r zed sTo; STO13 (lm Sl) adnos to paBallîno

  T apapop - e e nbTsTdo Uo-foa; - aun, p uaa-Teiq.

  aI -nod asopoTLta -4Sa a9xadgd TSUtW uoq.TsodUioo q ui 00o01 iauuop inod B4UQSTTfns nra, p T ^ luQnb 'aun' aBnoa u0 'nqIlrqsTp 14uauototIun ITos I $, nb ao r, nbsnf do-al suup 7i2: sa' < IT INASTiuoo-ti a, p ua.oui not asTATp: 4ueu $ iuT; 't; Toe 4uaTpai2u, - uoTsuadsns ap li oog iauuop inod nva, pa ^ u-sIT; ns i, 4Tu2nb aun suep uoTsBadsTp ua SluU uoIolo ap aouassaT B a ^ Um 2vjp, T o0 4amoJleqT4DD to the anblozuaq apTIo, T' enbFli4To pToi2, j T 0001 ds'b8TuoTS9p nexa 9 z uoa: 7o ap a: uessu 0 ss a u-e2-ep 9S I anb! ozuf q apTo 9 8 anTTSlUo aUTPIvu:: s4uaTpg2ut, p s4uteAins sE4T4ufenb Ba sadL SBP JTCp ds siaedia.id '4UOS' jt4e 4uBTpia2uT a, p 2 $ 5 '(ITU q) P; GB 01 e aEIBia [Tn aunlp BSOP anbeqo suep' 4ueu.uoo 'Iaefonq ToA.ed a2esn - asnanbS UoTsuadsns Aun, p ## EQU1 ## where: ## EQU1 ##, ## STR1 ## el suep s sJIITn Oz sal 4uebeldeaB uB 'T4Toe 4uaTPriuT a, p Sm 01 a 2ta cg ap sesop sap uuuuuuoo sUaBI7duoo Sap t lqeq2aies

  uo'T dp eldd uo 'aanpaooad seuu el 4uenbtidde u -

6L ú y 8 Z L 9 Z

26 7843

  Active Ingredient Micronized 3 g Polysorbate 80 5 g Ethylparaben 2.5 g Propylparaben 0.17 g Water for injection, q.s. 1000 ml. All ingredients, except for the active ingredient, are dissolved in water and the solution is sterilized by filtration. To the sterile solution is added the sterilized active ingredient, finely divided by means of an air microniser, and the final suspension is poured into sterile vials and the vials are

hermetically closed.

  The composition thus prepared is effective for the treatment of a neoplastic condition at a dose of 1 ml

(1 M) three times a day.

  Composition "F" rectal and vaginal suppository 1000 suppositories, each weighing 2.5 g and containing mg of an active ingredient, are prepared from the following types and amounts of ingredients: Micronized active ingredient 1.5 g Propylene glycol 150 g

  Polyethylene glycol # 4000, q.s. 2500 g.

  The active ingredient is finely divided by means of an air micronizer and added to the propylene glycol, and the mixture is then passed through a colloid mill until a uniform dispersion is obtained. The polyethylene glycol is melted and the dispersion in the

  propylene glycol is added slowly with stirring.

  The suspension is poured into uncooled molds at C. The composition is allowed to cool and solidify, then it is removed from the mold and each suppository is wrapped in a thin metal sheet. These suppositories are inserted into the rectum or

  vagina for the treatment of a neoplastic condition.

  Composition "G" Intranasal suspension 1000 ml of a sterile aqueous suspension for intranasal instillation, containing 20 mg of an active ingredient per ml, are prepared from the following types and amounts of ingredients: Active Ingredient micronis 3.5 g Polysorbate 80 5 g Kylparaben 2.5 g Propylparaben 0.17 g

Deionized water, q.s. O1000 mi.

  All ingredients except the active ingredient are dissolved in water and the solution sterilized by filtration. To the sterile solution, add the sterilized active ingredient, finely divided by means of an air microniser, and fill sterile containers =

with the final suspension.

  The composition thus prepared is effective for the treatment of a neoplastic condition, by ins-tillation

  intranasal 0.2 to 0.5 ml one to four times daily.

  An active ingredient can also be in undiluted pure form, for local use on the dermis, by

  intranasal, pharyngolaryngeal, bronchial or oral route.

  Composition "H" Powder g of an active ingredient in roughly fragmented form are finely divided by means of an air micronizer. The micronized powder is placed in

a container of the shaker type.

  This composition is effective for the treatment of a neoplastic condition at localized points, by

  apply the powder one to four times a day.

  Composition "I" Oral powder 10 g of an active ingredient in roughly fragmented form are finely divided by means of a compound of the same composition. The composition of this invention is as follows: ## EQU1 ## aqiaedas' I, U. 'IoA' ú e z z z z z z z io io io io Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top Top. ge: uo 'I eideaxei ap saesToqo suoaTsodmoo sap? lueum-iouoo saaedid 'I auIvsvTUTIloi ap saTe, 4Tun sanT2olosod samao; his z Tlduax! where 4uaTpeI2uTI ap 2 og a 2 S ZS 'g South snssap-To sTsITn og sael; ueIdu1al ue'; Toe; uaTpPaiSuT, p 2m OS e SZ 'g ap sesop sap; ueuauoo salnsdo sap ealqulques uoS; ap aredead uo 'eaIToiapdo apoui au9x al quenbTIddp u 5 lnoó Jpd sToj aueqnb aun saelnsdeo xnep e aun, p alggio uoT%; siutip- -id' anbTseldopu uoTIoa; I aun, p auama; wIg ai inod seot; a; uos salnsdeo saO a'lIan4Tqq BlTu -el a pginsduoua a aTQ X unasTuooTur unp uaeou nee 9sTAIp 4ueuauTI qsa jToe.uaTp; gnuTj oz! o 4uBiTppaSuT a, p 2u O? iuueluoo alnsdeo anbaqo 'eao uoITa2rSTUTUpP inod saT; aed xnap ua aenp auTe2ji, ap salnsdeo oo0 amedgmd uo a @ np auTIui2 ap salnsdo *>, uoT; TsodUIoD 5l anop mud smTo; amnb F aun 2m O & ap uoT'pelujuT, î med 'anbTsedoau uoToaj; B2 aun, p uemaT eal anod aeooT; ia 4sa uoT4Tsodmoo aeaO IT F mflnasTuOUOTUE a, p uaKou n2% uaeauT sssTATp Juos apquams2a; Oa uaeaiaTssom2 axisoj snos ;; $ oie uaTpm2uT unp 2 O0 uoT; tnsuI uoTI-soddoo nof med sToj aamenb aun nea, p aImaA suep uoTsuedsns ue alpnod ap s4aqovs xneap no unp alsio uoT% aqsIuT pQ, l aqd anbTsuidopu uoaToaej aun, p BuaeaTem aI / nod sa: oo $ T; a ss saipnod saeO s-anSed ua asui qa 2m Oz ap sallanpTnipuT sasop binds to; sInTp his xsTuoo $ Tu ampnod 2q 'JTe? mnasTuo-luo úy8L 9Z uoT: uaAuT aluasead VI ap aopeo al suep -4uae-a uoT -idposap aouaso $ d el ap ainqoal QI i?, I ep euaoq, l ap D ;; dsaI V quo pua A Tnà sunoTIe4dipp ja sa; uu; xjA 'suoTkeo; jTpom salt anb Fos ap VA II' anpuae.uT 0Z umaLuaIqQnbl-axauoSie ap sg) Topad snq xns sno% 4uojsTes tnb saooIT; ja a salaAnou senbsieldopuTuQ suoIeaedaad sap qa aouoTI; ea n-eAnou anbTseIdoguT! uV anaoe a gU4snliT% a TO 9 pT3T9-9 e 'ITnb alqTsTA Duae = 8eau 4sa IT' apQogad Tnb ao sQadea5 99ei aITeinlio a9u2I -el ap pie9g;, l -9 him / Sid 01 ap --E (j aun ja ggú, d aui-inu aptoqdmgI aeIonI -el ap p1a3, I 'agq, I apiaodaoo spiod ap 23 / z ap esop -el e% ot ,, nbsnp 1uellu and the apuueaSuolo-d a guuop - uoI: vedpad B-' elnFIsuI 0 [iaDue3 IFuo $ 4e np saloOlo-d sap 4uesTs ssgXsae r: 4: uo ja alduaxal Bb juaia = iojuoo sapaudgad; i9 -puo I aut% sstItIo ap saTl4iun sanbi5oiosod sauog sag (lI = / 2r.-51 x S 'b = "5Sa) 98dd -1, TA UT auTQQQQQQQQQQQQQGHUHQWHUHQWHUHUHQWHUHUHQWHUHQWHUHQWHUHQWHUHUHH 'p'pg8sU; QI apoqduJI ap p g, l? -' algq, l ap laiSodioo spiod ep S7 / $ G'E-Gl'O ap esop QI F% ZO e gz ap aTA eI ep luama2uoload L 9 z ú

Claims (1)

  1.- Antineoplastic substance called rolliniasta-   tine 1 and having the following structural formula H - - -0 H   1CH3 2 ..- Antineoplastic substance according to claim 1, having the following chemical NMR shifts I C: Oeca]. a9e Offset Offset Offset Chemical Chemical Chemical Offset #C (1C () C ()   1 174.5 s 15 74, Otd 25 32.8 + t 2 131.1 s 16 83.0 d 26 25.5 t 3 33.2 t 17 28.7 t 27)) 29/6 t 4 69, 9 d 18 27.8 t 31 37.4 t 19 81.1 d 32 31.9 t 6 26, 0 t 20 81O, 0 d 33 22.6 t) 7 29.5 21 27.8.t 34 14 , 1 q 12 29,3 22 28,4 t 35 151,7 d 13 25,7 t 23 83/0 d 36 77,9 d 14 34/1 + t 24 71,8td 37 19,1 q