Some Biological Effects of Vincaleukoblastine, an Alkaloid inVinca roseaLinn in Patients with Malignant Disease*

O. H. WARWICK,J. M. M. DARTE,ANDT. C. BROWNIN COLLABORATIONWITHC. T. BEER,J. H. CCTTS,ANDR. L. XOBLE|

(The Ontario Cancer Institute and Princess Margaret Hospital, Toronto, Ontario)

SUMMARY

The treatment of 22 patients with Vincaleukoblastine is described. The main toxiceffects were noted at the sites of injection, on the blood, and on the nervous system.Severe granulocytopenia with little or no depression of platelets was noted in nearlyall patients. In fifteen of the 22 patients the administration of Vincaleukoblastine hadmeasurable effects on the malignant process. Partial remissions of short duration,with marked improvement in general clinical condition, were obtained in three children with acute leukemia, in one patient with lymphosarcoma, and in one with breastcancer.

It has been reported by Cutts, Beer, and Xoble(2) that extracts of the plant Vinca rosea, wheninjected into rats, led to a marked fall in circulating leukocytes and to a depression of the bonemarrow. Subsequently a new alkaloid, Vincaleukoblastine (VLB), was isolated (8) from such extracts; it caused marked hemopoietic effects (1)and affected the growth of experimental tumors(9).

The chemical and physical evidence indicatedthat VLB had an empirical formula C46H58OgN4and was a member of a new class of dimericalkaloids which contained both indol and dihy-droindol moieties (4, 7). Rats given injections of0.35 mg/kg of VLB developed a severe granulocytopenia which lasted from 3 to 5 days. Lymphocytes were only slightly reduced and plateletsrelatively unaffected. Extensive depression, particularly of the myeloid elements of the bonemarrow, was also noted. Recovery of the hema-topoietic system occurred rapidly. In such casestoxic symptoms were minor, consisting of slighttemporary weight loss. No intestinal lesions developed even after massive dosage. Folie andfolinic acid, thymidine, cytidine, guanine, and

* This investigation was supported by grants from the Ontario Cancer Treatment and Research Foundation.

t The Collip Medical Research Laboratory, University ofWestern Ontario, London, Canada.

Received for publication January 11, 1960.

vitamin Bi2 did not prevent the hematologicaleffects.

VLB has also be reported (9) to have carcino-static activity. This has been demonstrated withL1210 and P-1534 leukemia transplanted in BDFhybrid mice, AKr leukemia transplanted in AKmice, and Sarcoma 180 in Swiss mice. In myeloidleukemia 1RC741 in Fischer rats, survival timewas not significantly increased, but a pronouncedalteration of the leukemic process in peripheralblood was observed. More extensive observationson the effects of VLB on experimental tumorshave been described in the preceding paper (3).In preliminary reports other workers have shownsimilar carcinostatic effects on experimental tumors (6) and human leukemia (5).

In view of the above experimental findingsVLB has been administered to humans sufferingfrom advanced malignant disease, at the PrincessMargaret Hospital in Toronto.

MATERIALS AND METHODSThe following standards were used in selecting

cases for treatment : (a) established forms of treatment had already been used and were consideredto have no further useful part in the managementof the case; (6) diagnosis had been establishedhistologically; and (c) physical signs and/or findings on special examination were such that objective measurement of any beneficial effects wouldbe possible. The 22 patients selected for treatment

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WARWICK et al.—Vincaleukóblastine in Human Cancer Patients 1033

included eight with leukemia, five with malignantIvmphoma, and nine with other malignant tumors(Table 1).

Apart from any effect which VLB might haveon malignant disease, we were at the onset of thisstudy equally concerned with any other of itsbiological effects. To this end the following protocol was observed when practical.

a) Daily hemoglobin, hematocrit, reticulocyte,and white blood cell count, platelet estimation,and differential count; marrow aspiration priorto and 1 week following initiation of treatment.

6) Daily weight, fluid intake, fluid output, andurinalysis: blood urea twice weekly; in the leuke-mias and lymphomas the blood uric acid was determined 3 times weekly in addition to dailytotal uric acid excretion.

c) Blood pressure immediately following treatment and twice daily; electrocardiogram beforeand after treatment.

d) Cephalin cholesterol flocculation, alkalinephosphatase, and bromsulfalein retention priorto, and 1 week following, treatment; serum elec-trophoresis by the starch gel method of Smithies(10) prior to, and 1 week following, treatment.

e) Photograph of malignant process before andafter treatment.

VLB sulfate was prepared in vials each containing 10 mg. of the crystalline salt.1 It wasdissolved in 10 ml. of distilled water prior toinjection. The dosage commonly employed in thestudy was 0.15 mg/kg, given intravenously for3 or 4 consecutive days. As noted in Table 1some patients received, after varying intervals,further injections of VLB in the same or increaseddosage.

RESULTSLocal effects.—Thefollowing effects were noted

in relation to the injection of VLB into antecubitalveins. Fairly severe pain was noted in one casewhen the material gained access to interstitialtissue. Three patients developed thrombophlebitis.In four cases numbness and tingling were notedin the antecubital and forearm areas, sometimesfor a week to 10 days following treatment and notnecessarily associated with the presence of phlebitis.

General effects.—The most constant systemiceffect was weight loss, noted in all cases. In someinstances this amounted to only a few pounds,but in other cases patients lost 8-12 pounds andin one case 16 pounds. Nausea occurred in four

1Prepared in the ColHp Laboratory, University of WesternOntario, from material kindly supplied by the Eli Lilly Co.,Indianapolis, Ind., U.S.A.

cases and was commonly at its worst the morningfollowing administration of VLB. One patienthad a brief period of vomiting following treatment,and another had diarrhea. Stomatitis and pharyngitis were seen only in association with markedleukopenia. Severe gastrointestinal bleeding occurred in two patients known previously to havepeptic ulcération.This was not associated withthrombocytopenia or prothrombin deficiency. Onefemale patient had temporary, partial epilation.

Symptoms referable to the nervous system werea feature in four of the cases. One patient hadparesthesia of the tongue and dizziness for abouta day. An odd behavior pattern unaccompaniedby abnormal neurological signs was a feature inthree cases. One patient seemed unable to understand spoken questions and walked along the corridor undressed. Another wandered about aimlessly and on two occasions urinated on his neighbor's bed. Disorientation and agitated depression

occurred in an elderly man in association withinfection and fever.

Hematological effects.—Asignificant fall in hemoglobin was noted in eighteen patients. Acuteleukemia was associated with progressive anemiabefore treatment, but even in patients with suchconditions as cancer of the breast, with an apparently normal marrow, the average drop following VLB was 2 gm. per cent. This was not associated with an increase of serum bilirubin or uro-bilin in urine. Depression of the reticulocytes wasusual, being most marked about 7-10 days following initiation of treatment. This was followed bya compensatory rise in reticulocytes and an increasing level of hemoglobin. In two patients amore severe drop in hemoglobin was related tobleeding from peptic ulcer.

In all patients a significant drop in the totalleukocyte count was observed within 24-48 hoursof the first injection of VLB. In twelve patientsa marked leukopenia developed, with counts below1,000 per cu. mm. and frequently 300-700 percu. mm. The maximum depression was notedbetween the 5th and 9th day following initialtreatment. In children this marked leukopeniawas invariably associated with agraiiulocytosis.Where the marrow was not primarily involvedby disease the agent affected mainly the granu-locytic series. Following the leukopenia there wasa rapid rise to levels above those noted prior tatreatment, as illustrated in Case No. 22 (Chart 1).

In 21 of the 22 patients VLB therapy had nosignificant effect on platelet counts even whenthrombocytopenia was present prior to treatment.In a patient with lymphosarcoma a fall in platelets

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WARWICK et al.—l'incaleiikoblastine in Human Cancer Patients1037

from 160,000 per cu. mm. to 60,000 per cu. mm.was observed.

In patients with leukemia the administrationof VLB was followed within 5-7 days by a phaseof hypoplasia in the marrow. At this stage anincreased percentage of smudge cells and otherdegenerate, atypical cells could be seen. Whenremission was obtained this hypoplastic phasewas followed initially by an erythroid and thena myeloid response (Figs. 5, 6, and 7). Whenremission was not obtained in leukemic patients,the hypoplastic marrow phase was followed immediately by reinfiltration with blast cells. Inpatients with carcinoma where the marrow wasnot involved by disease, a phase of hypoplasiawas noted within 5-7 days. This was rapidly followed by erythroid and myeloid hyperpla-sia. Throughout, the megakaryocytes showed nochange in appearance and no significant changein numbers.

Effect on other systems.—No significant change

was noted in fluid balance. Uric acid crystalswere noted following treatment in several patientswith leukemia and lymphoma, but there was noassociated hematuria, and there was no evidenceof urinary obstruction as indicated by oliguria.One patient with hydronephrosis and moderateuremia prior to treatment showed an increasein blood urea which quickly returned to normal.Another, with normal renal function, had a temporary rise of blood urea to 60 mg. per cent.In the lymphomas and leukemias a slight tomarked increase of uric acid excretion was noted.

No changes in pulse or blood pressure werenoted immediately following treatment or thereafter. No significant alteration was noted in electrocardiogram except for temporary changes associated with severe gastrointestinal bleeding intwo patients.

In several cases liver function was disturbedprior to treatment, as shown by cephalin cholesterol flocculation and increase in bromsulfaleinretention. The findings were not altered subsequently by VLB therapy. In one patient a negativecephalin cholesterol flocculation became positivefollowing treatment. No significant change wasnoted in serum electrophoretic pattern as a resultof treatment, although most patients showed someabnormality in pattern prior to treatment.

Effect on malignant disease.—It became ap

parent early in the study that VLB had, in somecases, a rapid and sometimes striking effect onmalignant disease. Fourteen of the 22 patientsshowed objective signs of improvement in theirdisease. This was characterized by decrease insize of lymph nodes, spleen, liver, and tumor

masses. In only five cases, however, could it besaid that treatment improved the clinical conditionof the patient.

In all the leukemia patients the spleen, whenpalpable, decreased in size during the course oftherapy. Also observed was a shrinkage in thesize of the liver and of enlarged lymph nodes.In three of the eight patients with acute leukemiapartial remissions were obtained. Accompanyingthe disappearance of blast cells from the peripheralblood was a decrease of blasts in the marrow, withsubsequent rise in hemoglobin and return of normal myeloid components in both marrow and peripheral blood. The remissions in these cases wereof 4 and 6 weeks' duration.

•••• ML.; ogt 45 Co of Breo«tV.L& OI5mq/K x4

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CHAKT 1.—Effect of VLB on peripheral blood of patientXo. 22—carcinoma of breast. Platelets were normal in number

and not affected by treatment.

Female, age 7 years, Case No. 6, leultosarcoma.—This patient

developed cervical lymphadenopathy 21 months prior to admission. Initially she was considered as having a localizedlymphosarcoma. Bone marrow was later shown to be involved.External radiation, aminopteriu (4-aminopteroylglutamicacid), prednisone, and 6-mercaptopurine had all given favorableremissions. Relapse of disease occurred 1 week prior to admission.

Physical examination revealed one small submental lymphnode measuring 3 X 1.5 cm. The liver was enlarged 7 cm. belowthe costal margin, and the spleen, 8 cm. Hemoglobin was 12.2gm. per cent; W.B.C., 3,950 per cu. mm. with 33 per cent blastforms; platelets, 140,000 per cu. mm. Marrow was hyperccllularwith almost complete replacement of normal elements by blastforms.

She received VLB 0.15 mg/kg on 3 consecutive days. Thechanges in peripheral blood are recorded on Chart 2. The sub-mental node was found to be smaller 2 days following the firstinjection, and 8 days following the initiation of therapy it was1 cm. in diameter. The liver was 4 cm. below the costal marginand the spleen 2 cm. At this time marrow was hypoplastic withan increase in erythroid elements.

The submental node, liver, and spleen became smaller overthe next a weeks. The marrow remained hypoplastic with a

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1038 Cancer Research Vol. 20, August 1960

relative increase of erythroid and myeloid elements. Blast cellsin the marrow fell to 32 per cent the 3d week after commencement of therapy.

The patient continued to feel well and was followed at weekly intervals. Six weeks following VLB therapy the liver andspleen again became larger, and blast cells reappeared in theperipheral blood. A second course of VLB therapy in the samedosage caused a drop in hemoglobin and in white cells. Thesubsequent rise in the white cell count was accompanied by theappearance of an increasing number of blast cells.

Of the patients with malignant lymphoma onehad Hoclgkin's disease, three lymphosarcoma, and

one giant follicular lymphoma. Following treatmenta change in physical signs was observed in fourpatients. In one case these changes were striking.

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CHART a.—Krfect of VLB on peripheral blood of patientNo. 6—leukosarcoma. Platelets 140,000 per cu. mm. prior to

treatment and not significantly affected by VLB.

Male, age 5G years, Case No. 11, lymphosarcoma.— In Janu

ary, 1959, this patient developed a swelling on the left side ofthe neck. Biopsy was diagnosed as lymphosarcoma. He wasreferred for radiotherapy March, 1959, and had a good response. Within 1 month, however, tumor masses had recurredin the treated areas and had extended to involve both sides ofneck, chest wall, and superior mediastinum. Further radiationproved ineffective, and he was admitted to hospital May 8thfor chemotherapy. Nitrogen mustard, nitrogen mustard combined with urethan, endoxan (N,N-bis[j3-chloroethyl]-N',O-

propylene phosphoric acid ester diamide) and prednisone givenover the next 3 weeks failed to halt the progressive course of hisdisease. Figure 1 shows the patient just prior to treatment withVLB. At this time he had a "tight feeling in neck," moderate

stridor and dysphagia, shooting pains in right arm referable tobrachial plexus involvement, and fullness of the abdomen.Free fluid could be demonstrated in the abdomen, and x-ray ofthe chest showed mediastinal involvement. Weight was 181Ib.; hemoglobin, 13.2 gm. per cent; W.B.C., 4,500 per cu. mm.;platelets, 272,000 per cu. mm.

On 3 consecutive days the patient received VLB, 0.15 mg/kg. On the 4tk day he felt definite improvement, and it wasnoted that the tense, blue appearance of his skin was less andthat areas of disease had diminished in size and extent. On days7,11,14,15, and 18 VLB was repeated in dosage of 0.18 mg/kg.

The white cell count was at its lowest (350 per cu. mm.) on the21st day.

With the larger doses of VLB the main systemiceffects notedwere nausea, starting 7-8 hours following treatment and usual

ly lasting through the following morning. On one occasion thepatient had diarrhea. Thrombophlebitis occurred in the ante-cubital vein of the left arm and was undoubtedly related toadministration of VLB.

One month after beginning treatment he weighed 165 Ibs.,and his general clinical condition was much improved. Allareas of disease showed marked regression. Nodes were nolonger palpable on the right side of the neck, and those on theleft side had almost disappeared. Crusting was present in previously ulcerated areas with weeping, healing dermis underneath. Hemoglobin was 8.8 gm. per cent; W.B.C., 1,250 per cu.mm.; platelets, 220,000 per cu. mm.

One week later there was a suggestion of exacerbation of hisdisease, and he received four additional doses of VLB (0.2 mg '

kg). Slight regression of disease was noted. One month later hewas readmitted with further extension of disease and with whatwas interpreted as a motor and sensory neuropathy. VLBtherapy had no beneficial effect, and he died 10 weeks followinginitial treatment with VLB.

Several features of interest were noted in theremaining group of patients with solid tumors.Case Xo. 19, with numerous subcutaneous métastases from carcinoma of the lung, showed definiteregression in size of these metastatic nodes, butthere was no clinical improvement in his condition.An elderly gentleman of 91 years (Case No. 20),happy and alert but with massive skin carcinomaof the back, showed no obvious effect of treatmenton the disease and his condition deterioratedrapidly. There was marked leukopenia and evidence of infection as indicated by fever. He becamedepressed, agitated, and completely disoriented.Death followed 5 days after treatment. The patient with pulmonary métastasesof chorioepi-thelioma (Case Xo. 21) showed no clinical improvement following VLB therapj-, but for 1 weekthe urinary chorionic gonadotrophins were at alevel of approximately 3,000 international unitsdaily, whereas the level prior to treatment andsubsequently was in the range of 12,000-10,000international units. Xo definite change was notedin the x-ray appearance of the lungs.

One patient with carcinoma of the breast (CaseNo. 22) had a satisfactory remission of 6 weeks'

duration following VLB therapy. Her case demonstrated well both the adverse and beneficial effectsof treatment, here described in greater detail.

Female, age 45 years. Case No. 22, carcinoma of breast.—

This patient had left radical mastectomy for breast cancer,February, 1957. Postoperative radiotherapy was given. Oneyear later a metastatic node appeared in the left supraclavicu-lar fossa, and she again received radiotherapy. In July, 1958,multiple chest wall recurrences appeared. Radiotherapy at thistime and in September gave a poor response. Oophorectomywas performed September, 1958, with no response, and prednisone failed to halt the slow, progressive course of her disease. InMay, 1!)59, she developed swelling of the left arm with extension

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WARWICK et al.—Vincaleiikoblastine in Human Cancer Patients 1039

of disease to the supraelavieular area. Again there was no response to radiotherapy, and she was admitted July 20, 1959,for VLB therapy.

The patient was well nourished and of good color. Supraelavieular nodes were palpable. The whole ehest wall was involvedby disease, the skin being bluish-red in color, thickened, and indurated. Disease extended into the neck region, over the deltoid, and posteriorly to the posterior axillary line. An ulcer 4cm. in diameter was present over the manubriurn. Its base wasnecrotic and appeared to involve sternum. The left arm andhand were edematous (Fig. 3). Hemoglobin was 14 gm. percent; W.B.C., 7,600 per cu. mm., normal differential count;platelets, ¿-20,000per cu. mm.; chest x-ray normal.

On 4 consecutive days the patient received intravenous injections of VLB, 0.15 mg/kg. Nausea and vomiting were notedafter the first injection. For 4 days she was bothered by a burning sensation in the area of disease. By the 5th day erythemahad decreased, the ulcer was cleaner, and its base coveredwith granulation tissue.

At this time the patient developed an odd behavior patternunaccompanied by abnormal neurological signs. She seemedunable to understand spoken questions and walked along thecorridor undressed. She was not emotionally upset and gaveindication that she understood she was behaving unusually andexpressing herself poorly. The only intellectual deficit notedwas slowness in arithmetic. She was depressed for the next fewdays.

Five days after starting treatment she complained of "aching pains in all my bones." W.B.C.,4,000per cu. mm.; platelets

normal. The marrow at this time was markedly hypoplasticwith absence of granulocytes, but megakaryocytes were plentiful. Three days later, W.B.C, count was 400 per cu. mm.with absence of granulocytes; platelets unaffected. The ulceron chest wall was epithelializing rapidly and erythema of chestwall subsiding.

On the lath day the supraelavieular nodes were no longerpalpable, and induration and thickening of skin had disappeared. There was less swelling of the arm. She appearedpale and haggard and had short periods of not seeming able tounderstand what was going on around her.

Nineteen days following treatment she felt "like my oldself." W.B.C, count was 15,000 per cu. mm., with predomi

nance of young cells including occasional myelocytes and mye-loblasts. The marrow showed reactive hyperplasia.

The patient was discharged feeling well, apart from stiffnessin the left shoulder. There was no apparent intellectual deficit.The ulcer on chest wall had completely epithelialized. Erythema of chest wall was slight with no induration or thickening.Hemoglobin was 11.5 gm. per cent; W.B.C., 5,100 per cu. mm.with normal differential. No significant change in platelets wasnoted during the period of observation.

This remission was maintained for 6 weeks. She has sincebeen placed on a regimen of androgen and thioTEPA (triethy-lene thiophosphoramide) with negligible response.

Post-mortem examination.—Autopsies wereperformed on eight patients, including three ofacute leukemia in childhood (Case Nos. 2, 3, and7), two of lymphosarcoma (Case Nos. 11 and 12),and one each of melanocarcinoma (Case No. 16),hronchogenic carcinoma (Case No. 19), and retino-blastoma (Case No. 15). Although the materialis limited, it is suggested that degenerative changesin lymphoid tissue and hypoplasia of bone marroware of much greater degree than is usually seenin untreated cases of these disorders. In all in

stances the marrow had undergone a profoundhypoplasia, and in three of the four lyinphoinacases a sharp reduction in the amount of lymphoidinfiltration had occurred. In the three leukemiaand two lymphosarcoma patients the number oflymphoid cells in the nodes was reduced. Degenerative changes in the individual cells included fragmentation and reduction in the amount of nuclearchromatin and a shrinkage and raggedness ofthe cytoplasm. Sometimes hyperplasia of the re-ticulum cells in the background could be seen,and fragments of nuclear debris were evidentin their plump eosinophilic cytoplasm. In contrast,no effect on carcinomatous tissues was evidentin the two cases examined.

Where viscera were involved by lyinphoma,changes were the same as those seen in lymphnodes. In Case No. 11 with clinical evidence ofinvolvement of the nervous system, there wasextensive infiltration of Virchow-Robin spaces bylymphoina cells.

No overt toxic effects attributable to VLBcould be found in any organs. No unusual ulcération or hemorrhage was found in the gastrointestinal tract.

DISCUSSIONBecause only a small number of patients have

been treated and essentially one dosage scheduleapplied, it is not possible to reach any final conclusions concerning the efficacy of VLB in thetreatment of malignant disease. However, it hasbeen clearly demonstrated that the agent has adefinite, though brief, effect on malignant disease.Of interest is the observation that the frequentlyobserved decrease in size of tumor masses andinvolved organs, such as spleen and liver, is seldomaccompanied by subjective improvement. Therewas a suggestion in two cases at least that, whilethe malignant process was initially sensitive toVLB, it later was unaffected by the same or higherdoses of the material. This development of resistance may be the same as that seen with othercarcinostatic agents. Other agents had been usedpreviously in our cases, and it would seem thatcross-tolerance is not a factor of importance.

It remains to be determined whether a betterinitial effect and longer remissions can be obtainedby a different dosage schedule and maintenancetherapy. At Dr. Noble's suggestion we are now

assessing the efficacy of a total dose of 0.2 mg/kggiven in ten injections at hourly intervals.

Of the toxic manifestations of VLB the bloodchanges are the most striking. The drop in hemoglobin in some patients following administration

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1040 Cancer Research Vol. 20, August 1960

seems more rapid than might be accounted forby the simple cessation of red cell production.Further investigation is indicated to determinewhether or not VLB has a specific hemolytic effect.

Granulocytopenia occurs rapidly, but recoveryis equally rapid and complete. Although the opportunity presented itself, we did not feel justified in treating patient with chronic myeloge-nous leukemia. The most remarkable feature ofVLB is that megakaryocytes and platelets arespared. Thrombocytopenia, even when presentbefore treatment, does not seem to be made worseby VLB.

The mental changes observed in several patientspresented a bizarre pattern with which we areunfamiliar. That it occurred in adults and notin children may be a coincidence.

Several observations would seem to indicatethat VLB has a mode of action different fromthose of other agents now used in the chemotherapy of malignant disease. Patients resistant toalkylating agents, metabolic antagonists, and cor-ticosteroids may répondto VLB. The patternof systemic reactions is unlike that which followstreatment with these other agents, and the relativelack of toxicity for megakaryocytes and plateletsis unique.

REFERENCES1. CCTTS,J. H. Biological Effects of Extracts of Vinca rosea,

Proc. Am. Assoc. Cancer Research, 2:289, 1958.2. CUTTS,J. H.; BEER, C. T.; and NOBLE,R. L. Effects on

Hematopoeisis in Rats of Extracts of Vinca rosea. Rev.Cañad.Biol., 16:476, 1957.

8. — . Some Biological Properties of Vincaleukoblastine,an Alkaloid Present in Vinca rosea Linn, with SpecialReference to Its Anti-tumor Action. Cancer Research, 20:1023-31, 1960.

4. GORMAN,M.; NEUSS,N.; and SVOBODA,G. H. Vinca Alkaloids. IV. Structural Features of Leurosine and Vincaleukoblastine, Representatives of a New Type of Indole-Indoleine Alkaloids. J. Am. Chem. Soc., 81:4745-46, 1959.

5. MODES,M. E.; ROHN,R. J.; and BOND,W. Effects of aPlant Alkaloid, Vincaleukoblastine, in Humans. J. Lab. &Clin. Med., 64:826-27, 1959.

6. JOHNSON,I. S.; WBIGHT,H. F.; and SVOBODA,G. H. Experimental Basis for Clinical Evaluation of Anti-tumourPrinciples Derived from Vinca rosea Linn. J. Lab. & Clin.Med., 64:880, 1959.

7. NEUSS,N.; GORMAN,M.; SVOBODA,G. H.; MACIAK,G.;and BEER, C. T. Vinca Alkaloids. III. Characterization ofLeurosine and Vincaleukoblastine, New Alkaloids fromVinca rosea Linn. 3. Am. Chem. Soc., 81:4754-55, 1959.

8. NOBLE,R. L.; BEER, C. T.; and CUTTS,J. H. Role ofChance Observations in Chemotherapy: Vinca rosea. Ann.N.Y. Acad. Sc., 76:882-94, 1958.

9. . Further Biological Activities of Vincaleukoblastine—an Alkaloid Isolated from Vinca rosea (L). Bio-chem. Pharmacol., 1:347-48, 1958.

10. SMITHIES,O. An Improved Procedure for Starch-Gel Elec-trophoresis (Vertical Starch-Gel Electrophoresis). Bio-chem. J., 71:585-87, 1959.

FIG. 1.—Case No. 11. Lymphosarcoma involving chestwall, neck, and mediastinum. Before treatment with VLB.

FIG. 2.—Onemonth following treatment with VLB. Chestx-ray at this time showed decrease in size of mediastinal masscorresponding to improvement in chest wall and neck disease.Healing ulcers show as dark, crusted areas.

FIG. 3.—CaseNo. 2-2.Recurrent breast cancer of chest wallwith ulcérationand extensive involvement of skin.

FIG. 4.—Threeweeks following treatment with VLB. Malig

nant ulcer has healed, and total area of skin involvement ismarkedly improved.

FIG. 5.—CaseNo. 6. Leukosarcoma. Marrow before treatment with VLB showing replacement of normal elements byblast forms.

FIG. 6.—Eight days following treatment, the marrow ishypoplastic and shows predominance of erythyroid elements.

FIG. 7.—Fourteen days following treatment, normal mye-loid elements have appeared in marrow.

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•*

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1960;20:1032-1040. Cancer Res   O. H. Warwick, J. M. M. Darte, T. C. Brown, et al.  

Linn in Patients with Malignant DiseaseVinca roseaSome Biological Effects of Vincaleukoblastine, an Alkaloid in

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