EP0159343A1 - Human calcitonin-gene-related-peptide - Google Patents

Human calcitonin-gene-related-peptide

Info

Publication number
EP0159343A1
EP0159343A1 EP84903749A EP84903749A EP0159343A1 EP 0159343 A1 EP0159343 A1 EP 0159343A1 EP 84903749 A EP84903749 A EP 84903749A EP 84903749 A EP84903749 A EP 84903749A EP 0159343 A1 EP0159343 A1 EP 0159343A1
Authority
EP
European Patent Office
Prior art keywords
pharmaceutically acceptable
peptide
cgrp
peptide according
acceptable salt
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP84903749A
Other languages
German (de)
English (en)
French (fr)
Inventor
Iain Macintyre
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Novartis Pharma GmbH
Sandoz AG
Original Assignee
Sandoz Erfindungen Verwaltungs GmbH
Sandoz AG
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Sandoz Erfindungen Verwaltungs GmbH, Sandoz AG filed Critical Sandoz Erfindungen Verwaltungs GmbH
Publication of EP0159343A1 publication Critical patent/EP0159343A1/en
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/57527—Calcitonin gene related peptide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to novel peptides, in particular to human calcitonin-gene-related-peptide, to processes for their production and to their pharmaceutical use.
  • the calcitonins comprise a class of peptide hormone occurring in a wide variety of chordate species including mammals. Though structurally related the calcitonins exhibit species related peptide sequence variation. In mammals, including humans, calcitonins are secreted from the thyroid gland, being produced by the thyroidal C-cells. Calcitonins play a major role in the maintai nance of body calcium balance.
  • rat calcitonin-gene-related-peptide or r-CGRP.
  • the existence of this putative peptide was predicted on the basis of nucleotide sequence data and corroborative evidence for its presence has since been provided by immunofluorescent localisation and immunoassay technique [Amara et al., Nature 298, 240-244 (1982); Rosenfeld et al., Nature 304, 129-135 (1983), and Fisher et al., Nature 305, 534-536 (1983)].
  • r-CGRP has been ascribed a sequence comprising 37 amino acid residues , with a surmised di sulphide bridge between -Cys- residues at positions 2 and 7 and a surmised C-terminal amide group.
  • h-CGRP human calcitonin-gene-related-peptide
  • h-CGRP is not found exclusively in neural tissue (as had previously been postul ated for r-CGRP) but is also present peripheral ly, for example in normal human thyroi d tissue.
  • h-CGRP as determined in accordance wi th the present invention is that of formul a I :
  • Peptides in accordance with the invention comprise not only h-CGRP itself (i.e. the compound of formula I in which the illustrated peptide sequence is unsubstituted at the N-terminal and ami dated at the C-terminal), but also other natural or synthetic precursor peptides which may be susceptible to cleavage under physiological conditions to yield h-CGRP, including e.g. synthetic h-CGRP pro-drug forms.
  • the present invention provides the peptide h-CGRP of formula I as illustrated above or a pharmaceutically acceptable salt or pharmaceutically acceptable complex thereof.
  • Suitable pharmaceutically acceptable salts of the peptides of the invention are e.g. pharmaceutically acceptable acid addition salts.
  • Such pharmaceutically acceptable acid addition salts may for example be formed e.g. with organic, polymeric or inorganic acids.
  • Such acid addition salt forms include e.g. hydrochlorides and acetates.
  • complexes are to be understood those complexes of known type formed with peptides on addition of inorganic substances, e.g. inorganic salts or hydroxides, such as a calcium and zinc salts, and/or addition of polymeric organic substances, and which are physiologically tolerable at desired dosage levels.
  • inorganic substances e.g. inorganic salts or hydroxides, such as a calcium and zinc salts, and/or addition of polymeric organic substances, and which are physiologically tolerable at desired dosage levels.
  • the present invention provides a peptide as defined above, in particular h-CGRP of formula I as illustrated above: a) in isolated or purified form; b) in pure or substantially pure form; c) in a form suitable for pharmaceutical use or; d) in characterised form; e) in extracorporal form;
  • isolated as used above is meant separated from other substances, in particular other chemical entities, with which the peptide is otherwise associated in nature.
  • purified is meant subjected to techniques of purification e.g. of chemical or physical purification.
  • the term will accordingly be understood as applying both to peptides isolated from natural sources and which have been subjected to a measure of purification, as well as to peptides obtained artificially, e.g. synthetically, e.g. obtained by techniques of chemical synthesis or genetic engineering, and which have been subjected to a measure of purification.
  • pure or substantially pure is meant having a purity of at least 50 % or more.
  • suitable for use as a pharmaceutical is meant in a form and, in particular, degree of purity appropriate for pharmaceutical application, e.g. in accordance with the methods hereinafter defined.
  • characterised is meant subjected to structural identification or to control of structural identity, including for example degree of chemical purity, e.g. for the purposes of pharmaceutical application.
  • extra corporal is meant not contained within the body or within any bodily system, e.g. within any organ, tissue or bodily fluid or within any cell or functional sub-cellular unit.
  • the term will accordingly be understood as applicable both to peptides obtained from natural sources as well as to peptides which have been obtained artificially, e.g. obtained by techniques of chemical synthesis or genetic engineering.
  • the peptides of the present invention may be prepared by any of the methods known and commonly employed in the art. For example they may be prepared by partial or complete chemical synthesis (e.g. by the solid phase method of Merrifield [J. Am. Chem. Soc. 85, 2149 (1963) and Advances in Enzymology 32, 221 (1969)], for example as hereinafter described), or by genetic engineering technique (i.e. recombinant DNA technique). Such techniques are entirely conventional and the present invention accordingly also provides:
  • a peptide comprising the amino acid sequence of h-CGRP, in particular the peptide h-CGRP of formula I above, bound to a polymeric resin support, said peptide being in free or protected form, to cleavage so as to free said peptide from said support, and, when required, carrying out process step a).
  • the following example 1 describes the method by which h-CGRP was first isolated and purified and by which direct structural analysis was first effected.
  • the following example 2 is illustrative of the methods for the production of the peptides of the invention.
  • This peptide comprises the C-terminal decapeptide unit of the r-CGRP sequence predicted by Amara (see above) with an additional N-terminal Tyrosine residue to permit linkge and iodination for radioimmunoassay.
  • the formula II peptide was conjugated with ovalbumin and anti-sera raised in rabbits by multiple s.c. injection of the conjugate emulsified in Freund's adjuvant (Difco).
  • Anti-serum specific to the formula II peptide was employed in accordance with regular radio-immunoassay technique to screen for the presence of immuno logic ally related peptide material in extracts of human medullary thyroid carcinoma (MTC) tissue.
  • Cross-reactive (i.e. r-CGRP- like) peptide was found in the extract of all seven human MTC tissue extracts tested. Cross-reactivity with r-CGRP permitted identification of this peptide as h-CGRP.
  • h-CGRP cross-reactive material
  • MTC tissue (15 g) was extracted by homogenisation in 15 % trifluoroacetic acid (TFA), 5 % formic acid, 1 % NaCl , 1M HC1, (Bennett et al., Biochem. J., 175, 1138-1141 (1978)), followed by adsorption to C18 Sep-Pak cartridges (Waters Associates, Hertford, Cheshire, U.K.), and then elution with 80 % methanol, 1 % TFA. The eluate was diluted 1:4 with distilled water, shell frozen and lyophilized.
  • TFA trifluoroacetic acid
  • the lyophilized extract was reconstituted in 30 ml of 30 % acetic acid, 0.01M ⁇ -mercaptoethanol and was pumped onto a SP-Sephadex C-25 column (83 x 1.5cm). A linear gradient of NaCl 0-1. OM in 30 % acetic acid was used for elution. The flow rate was 10 ml/hr and fractions were collected at 30 minute intervals. A small aliquot (5 ⁇ l) was taken from each fraction, diluted and presence of calcitonin, katacalcin and h-CGRP confirmed by radioimmunoassay.
  • Figure 1 is a representation of the obtained chromatogram.
  • h-CGRP eluted as a singe peak at fractions 62, 63, 64 (Fig. 1).
  • Fractions 63, 64 were selected for further purification on HPLC. These two fractions (CGRP content: 285 jug) were pooled, divided into 3 portions (2 ml, 4 ml, 4ml) and lyophilized. Each lyophilized portion was reconstituted in 5 ml of 40 % methanol, 0.2 % TFA and pumped onto a Spherisorb 5 ⁇ ODS (Queensferry, Clwyd, U.K.) Column (10 x 0.46 cm).
  • the purified material from two HPLC column was used for direct structural analysis of the peptide and sequence determination by protein chemical and mass spectrometric techniques, in particular employing FAB mapping strategy [Morris et al. Biochem. biophys. Res. Commun. 101, 623-631 (1981); Nature 304, 643-645 (1982); Biochem. biophys. Res. Commun. 117, 299-305 (1983) and Biomed. Mass Spectrom. 8, 463-473 (1981); and Barber et al. Chem. Commun.325-327 (1981)].
  • h-CGRP differs materially from the published predicted sequence for r-CGRP, in particular at positions 1, 3, 25 and 35.
  • the surmised presence of an -S-S- bridge between -Cys- residues at positions 2 and 7 and the surmised presence of a C-terminal amide group is proven and rigorously established for the first time for h-CGRP.
  • h-CGRP is prepared employing solid-phase technique.
  • Boc-phenylalanyl-benzhydril-copolystyrene-divynylbenzene resin (2.1g, 0.5 mmol) is subjected to the following cycle of washes and reactions: (1) methylene chloride; (2) 50 % trifluoroacetic acid and 5 % ethanedithiol in methylene chloride; (3) methylene chloride; (4) 33% dioxane in methylene chloride; (5) 5% diisopropylamine in methylene chloride; (6) methylene chloride; (7) solution of preformed symmetrical anhydride of Boc-alanine (1.5 mrnol) in 20 % N,N-dimethylformamide in methylene chloride; (8) methylene chloride; (9) 33 % ethanol in methylene chloride.
  • step (7) Each step is repeated as many times as necessary to ensure complete reaction or complete displacement of the previous reagent from the resin. This cycle of reactions is repeated for all Bocamino acids required to provide the sequence of formula I, except in the case of Boc-asparagine which is coupled in step (7) as the preformed 1-hydroxybenzotriazole ester in DMF.
  • the Boc-group is used for protection of the alpha ami no-groups of all amino acids and the following groups are used for protection of side-chain functional groups: Lys, 2-chlorobenzyl; Ser, benzyl; Thr, benzyl; Arg, tosyl; His, tosyl; Cys, 4-methoxybenzyl; Asp, benzyl.
  • the resin is washed with ethanol and throughly dried.
  • a portion (2 g) of the peptide resin is treated with anisole (3 ml) and hydrogen fluoride (15 ml) at -20oC for 20 minutes and at 0oC for 30 minutes and the HF removed under reduced pressure at 0oC.
  • the residue is washed with ethylether and extracted with 10 % acetic acid in water.
  • This solution is diluted with degassed water to a volume of 2 1, the pH adjusted to 6.8 with ammonium hydroxide solution and potassium ferricyanide (3 g in 11 of water) is added dropwise until the yellow color persists for 15 minutes.
  • the pH is adjusted to 5.0 with acetic acid and the solution filtered through a column of weakly basic anion exchange resin (AG3-X2, BioRad Laboratories), through a column of weakly acidic cation exchange resin (Bio Rex 70, acid form, BioRad Laboratories) and washed with water and 5 % acetic acid in water.
  • the peptide is eluted from the Bio Rex column with 50 % acetic acid in water, the solution is concentrated under reduced pressure, taken-up in water and lyophilized.
  • the peptide is purified by gel -permeation chromatography through a column of Sephadex G25 (Pharmacia Fine Chemicals) and by preparative HPLC on reversed phase as described, for example, by Rivier et al., J. Chromato. 288, 303 (1984). The chromatographic fractions are monitored by HPLC, and fractions showing sufficient purity were pooled and lyophilized.
  • the peptide was ca. 98.8 % pure as determined by HPLC.
  • the peptides of the invention in particular the peptide h-CGRP, and their pharmaceutically acceptable acid addition salts and pharmaceutically acceptable comlexes exhibit valuable pharmacological activity and are accordingly useful as pharmaceuticals, e.g. for therapeutic use.
  • they show potent vasodilator activity as may be shown in animal test models for example as follows:
  • Test substance e.g. h-CGRP
  • a solution of 133 ⁇ e 5-10 ⁇ Ci/injection
  • Intradermal injections are given in random block order according to a predetermined balanced site pattern, with 6 replicates for each of 7 treatments/ animal.
  • h-CGRP doses are tested against standard doses of a known potent vasodilator, prostaglandin E 2 (PGE 2 ). and phosphate buffered saline controls.
  • peptides in accordance with the invention significantly increase blood flow in comparison with controls, in a dose-dependent manner at dosages of from 25 f mol. and e.g. up to 2.5 p mol .
  • Observed potencies for h-CGRP are of the same or similar order to those observed for PGE2.
  • No increase in vascul ar permeabi l ity is observed using h-CGRP at doses of up to 2.5 p mol . as evi denced by use of i ntravenous 125 l-albumin to measure microvascul ar leakage.
  • the peptides of the invention in particular h-CGRP, also exhibit a potent influence on heart beat rate and force of heart muscle contraction, as may be demonstrated in test models, e.g. as follows:
  • Isolated rat auricle is suspended from an isometric transducer (Lectromed UF1) in oxygenated Tyrodes solution in a 10ml organ bath at 37oC.
  • the resting tension of the preparation is adjusted to 1 g and the tissue allowed to equilibrate for 90 mins. before commencement of the trial.
  • the rate of contraction is interpreted from the change in isometric tension by means of a Lectromed 4522 ratemeter and both rate and force of contraction are recorded on a Watanabe WR 3101 pen recorder.
  • Test substance e.g. h-CGRP, is added directly to the bath to give a final concentration of from 1 to 50 nM.
  • Propranolol is added to the bath to a concentration of 7.7x10 -6 M. This concentration gives a reduction in rate of contraction of ca.40 beats/min.
  • the test substance is added when the preparation has stabilized at the new rate (ca. 90 sees, after propranolol addition).
  • Peptides in accordance with the present invention in particular h-CGRP, produce an increase in rate and force of contraction in the above test method at concentrations of ca. 1 to 25 nM. These effects are reduced by propranolol.
  • peptides in accordance with the invention are observed to increase coronary flow as well as the rate and force of ventricular contraction at dosages of from 10 to 500 p mol, given as a bolus injection into the perfusate.
  • the peptides of the invention are accordingly useful as pharmaceuticals, e.g. as cardiovascular agents, in particular for the treatment of peripheral vascular disease (e.g. for the prophylaxis or treatment of disturbances in peripheral circulation, e.g. in limbs, including intermittent claudication) as well as for the treatment of coronary vessel disease (e.g. for the prophylaxis or treatment of coronary insufficiency).
  • a further area of application is in patient testing, e.g. in determining tone of the coronary arteries, e.g. in relation to application of other drug therapy or coronary surgery, e.g. coronary by-pass therapy.
  • dosages employed will of course vary depending on the particular compound employed, the mode of administration, the condition to be treated and the effect desired. However satisfactory results are in general obtained on administration, e.g. of h-CGRP, at a daily dosage of from about 0.0015 to about 0.15, preferably from about 0.0075 to about 0.075, and most preferably about 0.015 mg/kg animal body weight conveniently administered, e.g. i.v..
  • a daily dosage is in the range of from about 0.1 to about 10.0, more preferably from about 0.5 to about 5.0, and most preferably about 1.0 mg, administered, e.g. i.v.
  • Ix daily or in divided doses 2-4x daily or in sustained release form, and unit dosage forms for i.v. or s.c. administration suitably comprise from about 0.025 to about 10, more preferably from about 0.0125 to about 5.0, most preferably about 0.25mg peptide, e.g. h-CGRP, or pharmaceutically acceptable salt or pharmaceutically acceptable complex thereof admixed with an appropriate pharmaceutically acceptable diluent or carrier therefor.
  • compositions for use in accordance with the method of the invention may be prepared in accordance with standard galenical techniques known in the art for the formulation of peptide preparation, e.g. for administration by injection or nasally.
  • compositions for injection e.g. i.v., suitably comprise active ingredient, e.g. h-CGRP, in solution or in suspension in an appropriate, e.g. aqueous medium.
  • the present invention further provides:
  • a pharmaceutical composition comprising a peptide in accordance with the present invention, for example h-CGRP, as anywhere hereinbefore defined, or a pharamceutically acceptable salt or pharmaceutically acceptable complex thereof, together with a pharmaceutically acceptable diluent or carrier therefor;
  • a method of treating cardiovascular disease in a subject in need of such treatment comprises administering to said subject an effective amount of a peptide in accordance with the present invention, for example h-CGRP, as anywhere hereinbefore defined, or a pharmaceutically acceptable salt or pharmaceutically acceptable complex thereof; as well as
  • a peptide in accordance with the present invention for example h-CGRP, as anywhere hereinbefore defined, or a pharmaceutically acceptable salt or pharmaceutically acceptable complex thereof, for use as a pharmaceutical, (e.g. having a degree of purity suitable for use as a pharmaceutical) for example for use in the treatment of cardiovascular disease e.g. as hereinbefore described.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Toxicology (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Endocrinology (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
EP84903749A 1983-10-12 1984-10-04 Human calcitonin-gene-related-peptide Withdrawn EP0159343A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB8327346 1983-10-12
GB838327346A GB8327346D0 (en) 1983-10-12 1983-10-12 Peptide

Publications (1)

Publication Number Publication Date
EP0159343A1 true EP0159343A1 (en) 1985-10-30

Family

ID=10550087

Family Applications (1)

Application Number Title Priority Date Filing Date
EP84903749A Withdrawn EP0159343A1 (en) 1983-10-12 1984-10-04 Human calcitonin-gene-related-peptide

Country Status (6)

Country Link
EP (1) EP0159343A1 (ja)
JP (1) JPS61500119A (ja)
AU (1) AU562318B2 (ja)
GB (1) GB8327346D0 (ja)
IT (1) IT1213287B (ja)
WO (1) WO1985001658A1 (ja)

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE134631T1 (de) * 1983-06-15 1985-12-05 Roger Kingdom Craig Peptide, pharmazeutische zusammensetzungen, gene, vektoren, wirtorganismen, verfahren zu deren herstellung und diagnostische reagenzien.
DE3685885D1 (de) * 1985-01-16 1992-08-13 Ciba Geigy Ag Oligopeptide sowie zwischenprodukte und verfahren zu ihrer herstellung.
HUT44794A (en) * 1985-12-04 1988-04-28 Sandoz Ag Process for production of calcitonine-derivatives and medical preparatives containing such compounds
JPS63126894A (ja) * 1986-11-17 1988-05-30 Toyo Jozo Co Ltd カルシトニン遺伝子関連ペプチド
GB8720115D0 (en) * 1987-08-26 1987-09-30 Cooper G J S Treatment of diabetes mellitus
US5266561A (en) * 1988-01-11 1993-11-30 Amylin Pharmaceuticals, Inc. Treatment of type 2 diabetes mellitus
US5175145A (en) * 1988-08-26 1992-12-29 Amylin Pharmaceuticals, Inc. Treatment of diabetes mellitus with amylin agonists
JPH02229119A (ja) * 1989-02-28 1990-09-11 Toyo Jozo Co Ltd 動脈硬化予防および治療剤
DE3943519A1 (de) * 1989-04-27 1991-01-17 Stief Georg Arzneimittel zur behandlung erektiler dysfunktionen
US8168592B2 (en) 2005-10-21 2012-05-01 Amgen Inc. CGRP peptide antagonists and conjugates

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO8501658A1 *

Also Published As

Publication number Publication date
IT1213287B (it) 1989-12-14
AU3503684A (en) 1985-05-07
GB8327346D0 (en) 1983-11-16
WO1985001658A1 (en) 1985-04-25
IT8448978A0 (it) 1984-10-09
JPS61500119A (ja) 1986-01-23
AU562318B2 (en) 1987-06-04

Similar Documents

Publication Publication Date Title
US5583108A (en) Vasonatrin peptide and analogs thereof
CA2071538C (en) Hpth (1-37) fragment, its production, drug containing it and its use
WO1990001940A1 (en) Atrial natriuretic peptide clearance inhibitors
EP0464105B1 (en) Melanin-concentrating hormones and dna for expressing the same
CA2100743A1 (en) Amylin antagonist peptides and uses therefor
AU562318B2 (en) Human calcitonin-gene-related-peptide
TWI230613B (en) Peptides having potassium channel opener activity
US3880826A (en) Vasoactive intestinal peptide
AU8237187A (en) Derivatives of atrial natriuretic peptides
US4721704A (en) Potent synthetic atrial peptide analogs
WO1985001284A1 (en) Synthetic peptides with egf like activity and pharmaceutical compositions and methods of use
Mayer et al. Potency of natural and synthetic canine gastrin-releasing decapeptide on canine antral muscle
CA2202496C (en) Neurotrophic peptides of activity dependent neurotrophic factor
JP3025672B2 (ja) 新規生理活性ペプチド及びその用途
JPH0570484A (ja) ペプチドおよびその塩
JP3264439B2 (ja) 新規生理活性ペプチド及びその用途
JP3258639B2 (ja) 新規生理活性ペプチド及びその用途
JPH0570482A (ja) ペプチドおよびその塩
JPH0578391A (ja) ペプチドおよびその塩
JP2883903B2 (ja) 新規生理活性ペプチド及びその用途
KR101130283B1 (ko) 평활근 이완활성을 지닌 칼시토닌 관련 펩타이드
JPH0570483A (ja) ペプチドおよびその塩
JPH0570492A (ja) ペプチドおよびその塩
JPH0570485A (ja) ペプチドおよびその塩
JPH0570487A (ja) ペプチドおよびその塩

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

AK Designated contracting states

Designated state(s): AT BE CH DE FR GB LI LU NL SE

17P Request for examination filed

Effective date: 19850816

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: SANDOZ-PATENT-GMBH

Owner name: SANDOZ AG

Owner name: SANDOZ-ERFINDUNGEN VERWALTUNGSGESELLSCHAFT M.B.H.

17Q First examination report despatched

Effective date: 19870204

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION HAS BEEN WITHDRAWN

18W Application withdrawn

Withdrawal date: 19871014

RIN1 Information on inventor provided before grant (corrected)

Inventor name: MACINTYRE, IAIN