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1. What is the absolute stereochemistry (Cahn-Inaold-PI'Clog nomenclahrre) about the chiral centre(s)

:::(o:e~::? S\:onOCMfr\\~-tvy ohut Cd = R

~J;«:) 'S-toy-(oc'0.QJY\\)~v:J Octp0-t C3 -: R.

./ I'll

HaP 3, ,CH2 (J'\

G- vi \L )

CHa

l-isoleucine

2

2. What is tile absolute stereochemistry (Cahn-Ingold-Prelog nomenclature) about the chiral centre(s) of L-threonine? (2 pts.)

~ - 1

HaW ........... 2 ,.......COO· qH

. CH,

H3C"""'" 3 I"OH

'S-le> ve 0 U'\..Q tq~>1 :)-\(~ S te v t cxJ'\.Q n'l \ ~~\ )~

a.bQLtt cJJQ0~l-

L -threonine

2

3. Where would yon expect residues like valine and isoleucine to be within the tertiary structure of a protein? How does their location within a protein affect the attractive forces between charges? In your brief explanation please refer to Coulomb's Law for full credit.

HaW, ,.......COO- ~Q.\. 1.k.JCA...,\\ 0., txFX(~ 5-tY'vlL:t:,ljr~ h\:JL \)0.\ \~

C.H c,..Y'Cl \ ')0 \ Lue i (,-CL.- -b=:> be.. l/\ -t'r\..(L. cJ:)1--c....,

HaC/CH'CHa of . tl'\JL 'ero-u l (\ T\r\Q-rL.. O~ (\onp-:Jlu1 S I C\.JL j rou~j <:) n -t\'\..QS L. Cd'Y\ 1(10 ac k::i.:J

(0") l-valine C\\~ W~ \,UOl.,\ \.l\. ~'(J._\f\t -tD fc{Cr OU:C .

'---' _G\.-l\ l 'JO\\JCV -t S SL\ Ch CL2:, \J,-(xtr\f. WO,--CQ( 00> \'\Q~ Q Yl \9 h

c\ \e. \ Q. C \. n c ccNl \{ elVI t 6rd Cc \;1 . -t \~QJ'f 'fuJft_ \~d!

C VV)\f C\C~ ~\P(U -t l,~oc\ l{\~ -t;o IO~(.r eM_ (9 '-l, of '. u~':;O() lA -t\ 01\ loct~ CQ Y\ cha; ejL C, PI (LO(ull/\~ to ~ c0u\of\f\b I~ \Q\}J C 1\ \1\\)( ()-{k-i Pi(Q1Cbv~()/\(-'\.. I k.'0

b-\ L\ l_Ct Vl C 0\J \Il 'S t-(A (\-t l D ')

.-- \ .

t. ==- f-.£l' q; / Dr

4

5. Let's say that the hydrolytic Enzyme X has a valine residue in its active site. The first order rate constant (kvat) with respect to substrate is 0.001 S-I. If a threonine residue replaces the valine residue, a new hydrogen bond between substrate and enzyme occurs that results in a 3.0 kcallmol favorable interaction resulting in a rate enhancement. Please tell me what the estimated rate enhancement for the reaction is, and then estimate the rate (kThr) for the new enzyme,

k \leI. \ ~ O. 00 \ S- \ bt:k} -+ -::: ~. 0 \c..coJI 1'1'\ Q \ -z: -:; 000 cc,,- \/n'lu \

\2-0. t t <-- f\ ~Cl Y\ CQ i'Y\. __ QJ).-t - ¥X -en (' K \jO" \

.', ~-thl ~ (\C;;~)62 ~, cclr cc[O~\5qs-\\

6. In the previous problem I made an assumption that the favorable binding interaction results in a rate enhancement. Please help me understand (briefly) why this assumption is likely to be false using your thorough understanding of Michaelis-Menton enzyme kinetics,

l -r S ~ cS ,:L~E '\- \=J

1::-1

-77



! Ir. t- d -+-in n /l '1\ a H tr\Q.... 'Sub )-tr(X-D1 I \ tv . I () c r LQ e Lf r <,

C<::fV\plQA stXD~;r -ttv> h kL \.0\ II w

\ {On \r\j -t~o ct 'S lC)UJ C r rT Q C { I () n ret {J

( n eye ((~ -to ( n t'""M n CJ /r.:« 1//-(- ')

L \.~O ~ l~l;-)l-t)

\ ,"60 0-10, os : (-)( -t'') (--\ ') \0, O~ - c\ :~3 ~ (- )(-\ ')(t-)

') c\.~:' :c-)l*'')li<-)

. H(\

N

.

H3N7 COO-

L

pK2 = 9.33 . pK1 '" 1.80

~~'J;

.; ;!L:histidine

~ (j2.)

\. 0 0"

H3NT = Hb

pK2=9.9 ~~-R histamine

7. What is the absolute configuration of the alpha carbon of L-histidine?

'f-\ b~O\V:-\"'L~ \uY\ -\l3L.tY' CI.. {jo n = (~')

8. When the pH = 7.0,90% of the imidazole side chain on L-histidine is uncharged. Determine the pKR far the imidazole sidechain .

.:/, \0\\'\ LCl-t \0 n -= \ of -= \ QO / ( I -\- \0 ('1,0 -pl)2 ) )

, +-- \O<-1.o-f\L~) z: 10

\ 0 -.:t.G·-f1't~ ~ 0 :. ~ ,0 -pr-R... ~ leg C1

~?K rL -=: (;~ oS7( .

9. What is the isoelectric point (PI) of L-histidinei You must complete problem 8 first to do this calculation.

10. The com ersion of t-histidine to histamine is catalyzed 9Y Histidine Decarboxylase (HDC), which is a pyridoxal phosphate (pLP)-dependent enzyme. The enzymatic decarboxylation ofLhistidine in -olves the stabilization of an unfa\ arable negative charge \ ith the PLP prosthetic group. What kind of covalent. catalysis is likely to be involved? '

11. Despite having au sp2-hybridized alpha carbon intermediate during HDC s decarboxylation orr. histidine, let s say that we've experimentally determined by NMR studies that the enzyme adds a second hydrogen stereospecifically to be re face of the Sp2 -hybridized alpha carbon to form histamine. Please indicate which hydrogen is the pro-R hydrogen atom (either H, or Hs).

·t-I~

1

~-

-,_I-._..:~

yh-= Li ,q ?Ll2-~ ~.o

6

./

1f·~at i? tbe overall charge on histamine at pH = 8,9?

'/, \of\\lo.tlf.)1\ of ft-2_::: qo,~ -1- (-\: c:.ha.(~-y

"{. ~ u{\1 t<>-"\\{I:'\ r.A - ~t\t '; O,j ;'1..1 . .0' [. l\~o.. ')

O\J (((:A \ \ po~ i -t'! V C L \r\4Y<j-~ .

13, What kind of inhibitor (competitive or uncompetitive) would you expect Goo fluoromethylhistidine to be for HDC?

Gorn\X-t'I-t'r...Jc _ .

.--- S\ fY\ \ \ 0..'( Soot V\J c-tJ_J.I~ C cu'\

'b \_"d -Co CVYZL(I'Y\s!...-/

o.-fluoromethylhistidine

14. What is the effect of c-fluoromethylhistidine on the enzyme's apparent Vmax and Kj\.j? (2 pts.)

c~~Or-(rrt o-pp&\rtn t

\J vn CA)L - r\O C \)0 \'l9L k-m ~ tnGrLCtSC

15. Please show using an Eadie-Hofstee plot (\loI[S] rersus v 0) the fo11O\ -ing: what would happen when no inhibitor (nfluoromethylhistidine) is added over several substrate concentrations; when [1] is twice the K\; when [I] is four-fold greater than the Kl- Label each axis on the graph, draw the corresponding lines, and indicate changes to the apparent max and K-M.

V0U:::' ')~AppoH d -00 _

ht. c'-- \-',-;--c,\~ rvl \ ,1"\ ( 0

7

16. Circle the pharmacopho . (3) . . .

nc groups on the structure of o-fluoromethylhistidiue below. (Spts.'

a-iluoromelhylhislidine

17. Describe briefly (on this 1 ) . .

i: h f h '1 1 page on y a way to test the relative Importance for biolozical activity

lor eac 0 t e..J p rarmacophoric 0 . f fl . . - i::>

(SAR)- di o10UpS 0 G- uoromethylhishdme for structure-activity relationship

stu ies? In your explanati 1 . .

_ _ . . . IOn pease provld5: some ratIOnale for the substitutions/ chancres that

)'Ol~ are suggestmg. -- - ~ - '" '-

r- a~ \.0Cl~ 'cO .J--,,_Qsk.. k'0JL C\l.\c\V\-t~ W( 'J'"\.L pluv (Y)o.CC{JhO(1 :ro~~ I:':, ko CAI-\:_F *VvL. jl:.fIH .. --t-Urt., SIl5 ht~ arc),

StL I-\ow ~ Qc-tI\.J\-\:"1_ChCtnrys- '0'f rLrlCtCl~

-~ ,pr,Gt(mc.lCorhGr"'~ wd:h 0.(\0\ ~ -that OK...-

cN-\Y\ \ CC.l\ 1'1 s\ rc. I \ C! V l<:OSY\Q_ ~d\Qf1ll t --f:-YLo. 0. C t-i \ft~ "S h a.M \'\o-t cY\o.~'19t . .J S \3\\ Ifl CQY\l ~ . c.\ e.- ex cho,~ u ~ ex: .fhAv f'r"ClCD 'f*'PI'I L. .3 YoLlp lAJ1th_ CA ~ - 10/0/ st§-( rt: ')

_/~Qt (OXI oJso ~)t B'~ vC\vlC(tlon a+ bio/Dj/cc{)

CA({\VI-tU w\{h c\\{ttl'1..ll){ \"--'ibs-u·tCL.e n{J bc{)(CL 0 n thQ IY h'j ~o b; ci-ltj «;y:j lr\QJJ--lfu.'-I I') Li I' -t/ ti Of) \ n cet 0(10 \

ov\cl \A..l6\U V.

, \'c\V1 0--1.'> ct: \'\Q v f' h,-\_'i LDch,Q m \ C: 0.,.1 ~y -tits 'Of -0{Nj

• i. ., • «1. rL n' ..J- <:l \\0 -+ .~ nkJ cu,J\ \-:J c..--

(Ylul e eli "-.fL u,J-t h cl ry\ I L..-> \....--\ S '- I , .

o i:J Kvvc:cl or0- cdY\\0",rc6 -I:D k:~ yxtnJ\-t mol c w LL

(Ski; vi c. -fCt~ ~o v;' - T or-t ' ~ 59 clCl-t1 0 (\.i c c~ vYlct a v re- i-r act / J: G-,J, C

-11'111\ \ r.C».J \_p-t vJO-A.\6 d,Qvtl9') Q \(~dfo(n1u~utr:n w \-t \f} 3-.\ \'YS -tl-CW n-tstro-l 1\Jj'\) VI Cl. L D!YO teo: \ CJ C -(: I Vi '-t I

'~'L\l (J~. (QS A K tQ C \"t \'1 '1 LA.Q \)

\ -'-~

-+7/1

-,

8

2-methyl-3-phenylpropanol (MPP) acetate

1

18. Please draw the S-enantiomer oflvTh1P-acetate. o

~I Wo~~

4

Ci:J (C/I

19. The apparent enantioselectivity of the lipase-catalyzed hydrolysis of (S)-IvIMP-acetate is 16-fold greater than the hydrolysis of the R-enantiomer. (Enantioselectivity can be defined as a ratio of catalytic efficiencies (kcatIKM)') If the kcat for the two enantiomers is rougbly the same, and the K.,vl(R) = 4 mM for the R-enantiomer, then what is the lipase's KM(~ for the S-enantiomer?

(J~r(iVK + I ~ '¥- ~ s k.\'t<'c;.;)l R.. ::; I-j (\1) m

(V) \ :: \\0:: /-Y

Vu; \em)\( . ~1\Q. Cc0

t \f\,') --=:: \ ~ ( ~ ,0 r(\ (YI ) -::: [ \0 '-\ 11, ''), 1

20. What is the free energy difference between the two MlvlP-acetate enantiomers (MG) given the

Enantioselectivity(S) :::: 16? ;:::.b.G + 1[2. T

\\o~-e

.'. (\ \~ \ 10 ](\ ,C\8"1- (C~ \ \C-~ \ )'\'\21\ -I )( d gg \( ') -;:- 6L~q r

~\, ~ CC,--\ 11,0\- \ (

21. If some key residues of the lipase were mutated so that the difference in free energies between mutant and wildtype proteins result in a 3.2 kcalfmol enhancement in binding to C.5)-lvllYIP-acetate then what would be the corresponding increase in enantioselectivity?

r-: '. \. ..J- _ C::'(j.c:ocu'vrnol')/(IP1J1-C.o..\t--'lY\d-')(l..CJJ tj

c'(\co. . .\'\ -\:\ O,}L ( C:-t\ v\ 'v~ -

22. The cause for enantioselectivity observed for MlVIP-heplalloate is found to be dependent on differences in kcalo whereas the cause for IVliVIP-acetate is dependent on differences in K;\>I. Are the differences between 1vlNIP-acetate and lVflVlP-heptanoate substrates due to geometric or stereochemical specificities? Choose one.

6](OfY\(tt~ 1JeCJh Ci -t,·i e)

~)

(0)

/ /"

9

Marks

x

:-:H~

a

! Capsaicin analogue

OH

Table. QSAR data on the capsaicin analogues

fcompound Ix Lo-.K (1/ECsot IT MR
1 I H I :1.07 I 0.00 1.03
I
2 I Cl -0.09 0.71 I 6.03
.... 1 N02 -0.66 -:0.28 7.36
.)
4 I CN /&,1. -1.42 . 0.57 6.33
5 I C6H5 10 " 0.62 1.96 25.36
-J;~Lf·
6 I N(CH3h -0.64 0.18 15.55
7 I I 0.46 1.12 13.94. 1

23. Which X substituent results in the most active capsaicin analogue?

C \ -r=Jr ('_0'~\.

~ I C~ lDl\'\you\,cl -ttl{) Jl~ «» \] )

(LCj~ ~.\ L1. Z I "3 2 L '1- ,-,=I')- .LiG

)-<-:,::,' 1.-1_ !:;L

24. Let's estimate the LogP of compound 1 (where X = H) to be 3.5. What would be the calculated 10gP (CLogP values for any hl/o compounds otber than compound I?

25. Molar Refractivity (kfR) is a measure of a substituent's volume, Which substituent occupies the largest volume?

1

26. According to Lipinski's "Rule of Five' > a CLogP > 5.0 is an indicator of poor absorption. compound will have the highest biological activity and an acceptable CLogP value?

(\--J l CO\'Y'f>Ou\'\c\. ~ y)

\Vhjch

27. Other than the methyl group on the methoxy substituent of the capsaicin analogue, only one other carbon is Sp3 -hybridized. Please suggest one change in the structure that would result in a reduction of free rotation of the molecule about_ that sp) -hybridized carbon.

- net o, Ct. dQ,-\.b~ )~I\o\

J

4

u I

-'

-:

10

(S.Bandiera) l'vlarks

28.(a) Briefly explain the purpose of drug metabolism/biotransformation in the body.

CO\'\ \I(,J -t'l (J,. \ I po r " i yc f'0.d e CJ.A l~ .lryt.Q. CJ... Yl'I0rc t\\.\ cXV"Or hi \ \ ~ rncl~'--~ such -t.,\'1D -t I i. co. n 1'J~

Z XL r-C-U d. f0v Vy\ -t ~ k:~DC11 V \ o; 0 Y' i (\,..Q. Q { {-( Ce..S -' e f r .

3

(b) Explain and discuss three possible outcomes of drug metabolism/biotransformation. You may use examples if you wish.

CD ~n CtC-t\ V c. dMC\ COS, DC COi\\J (y -t.Q el -to aVI -

\(\o.\-t\\J~!""--Q{ol6o\\--tp lYlG 1()~r9LV ha~ iphCtYVY\uco!Oji(4)

rAC~lv\~ \1\ -tk ~Joct~ J .

Q> 'An \(\ ,5YC-t.1 vc prac.\V\.lii Cc..Y1 be convU-D 0.. -c..o Cl..Vl C~C't.ive C\~ ()\.Jtiut06!\--tp lSCtll\\ pho\/rYlO-Co{aj,('c""l QrtIV\~ (\ -ths '6ccl~) - .

CI>f)V} _ ulti,,!L,; dvuo CC\.Yl bG CDf\\JCy-vct -Go G\. (y\.Jltqbof,-f.;J v \-t\0 ~6\~rin-f.. rhClVffiCcQ)[D,S icet! c..c-tiv;~ In t~ bCCt ~ C ncl,j Cl \'VJ..\) ---t Ct rg,ct :s

29. List two characteristic features of Phase n reactions that differ from Phase I reactions and describe these features as applied to Phase I and to" Phase II pathways, highlighting the

differences. .

?hC\\L ~- net l {( ons fLA..nett 01,\ -Lo Oc\c\ Qr'- eXectG D-- \"f\O\,---'7 \\y c;\~r \1 i.) \ "c j\'\JLtp <0 n ~ dllA.L\

f\'\a\e. cutQ-'s u)\,\\ G ?~'-'t,OL n '---LU C-ti Q -nj ~rYY\ cov(J,\Q Vyt - bOf\Oj V0 \ -t h hy Ci YO P \'\ \ I \ 'c cDnj ~)':r'~ 'Cp~ . o I"\:{ 0 -tl"'-Q._ c\ V\J.j nl C) \ ~-LGl \s .

'PnCt IG -1- . n Ctc:tw V1-l -b1 Ie! i ,o.\ kt 0 C( u y V ,. C!

(Y\ \ C'\"DS 0 YnCl \ /~- -}\'[ lj nLQ,j - e IrA j L JL i"--Co..(-t I 0 Vl..) -O.ce u.r l'Y\OI'J mqiG\.J! h~ ViCA ~tC),>OI'Y\Cl \

Q(YLLjn\Q_s (~orY~ eK(Qr?t i C7n) ) _. I'

Cy) 1I"O)'1\J 0( CLlS! on J Q 6\v1A.Q i'Y\O\ e.LL{ le c.,LA II LA~.ao 'g-o el'\G\I,L I rfGtctlOfJS -to b(YDn~ m\)\'e.. hycA~v)'ll lie

Ol,\(,l -~h,.Q ('\ l(r()Jl i'jO -),-,lb ~{~ U-~ r'rt pJ")ct ~'L II r-~ rLS -t'O 30\\(\ {\JC,\ m~)IC \'\\...'\ c\~)hl'\L~ t.Y (11'\0-{ OlWClL/(

-tNi CCl)G-j 'Son'U2 . cVqqJ (}, ,vI oVlly l,{VIClQi'g 0

,JI'\ct \G If- ~ n "--'t o~ ~ VI Y I

/

Marks

11

30. For each of the reactions shown, name the enzyme that catalyzes the reaction.

CI

1

(a)

1

(b)

1

Cc)

1

Cd)

.>

3

~OOH

H -b-C1

O--C---CH2 -

\/ \

~2 0 \ ;} C~2

COOH

Alcofenac

CI

HO

\ CH-GH2 / \

HO---CH2 0

-- ...

/-

Plt6&; (j:...O, hu. Olfb\U SG

'f I

Aspirin

12-

31. (a) For each of t .--'''Z,ing drugs, draw the strllctures of two possible metabolites that would be ~onned by tw dlfferent enzymes belonging to the pathway shown and (b) name the enzymes involved,

(i)

2

r,5

2

(ii)

0
H
2 H3C'-..,. )
N
I\,/ O~N
I
CH3 Phase I enzyme ..

N arne of enzyme

o

Phase I enzyme ....

Name of enzyme

C '-19 ()-J ~ eM 0..\ \(~i \ oA:) 0 f\ ')

Phase 1 enzvme ..

Name of enzyme

(~!~ (N~O)(lctO.-U0(\)

V 2

2

,

.:

r> [)

2

2

(iv)

COOH

OH

,

Nameofenzyre I

G \ ~w::r OS'-1

Phase II enzyme ~

Phase II enzvnie ~

f 1Z\I.me Name 0 eL .J .

G'UlY.fD ,~n~\ , . \,rCI\f\.S -\trO,)L

\ ~ "',

.,..-~.-

.!

3

..,

j

cl

I

4

14

32. Propran?lol.is a ~idely prescribed l3-adrenoceptor antagonist. In hu.mans, clear~nce via hepatic metabolism IS rapid, The pathways involved in propranolol's metabolism are outlined below.

+ I~r/

OH

..-7 HOOC

H~~O

(3)

(a) For each of the numbered pathw ays shown, giv e the name of the enzyme that catalyzes the reaction and list the required cofactors.

(1) ('\ ~ J(~v\)\"f\o.-t\ L \\\;\c\t"'\)f-~ \ cecil] (\ ') ',Y\ee.d,~ N'A uP-+-

(2)~\\-f0k0(d(~~C(o.~e : \':.\I;P

(3) "E"ALALUY''O(\O')L{i -tYCti'isfcrosc : Nf1 \,:yt J

(b Compare and contrast enzymes (2) and (3) in general terms (i.e. not relating specificaIIy to the metabolism of propranolol) in terms of their substrate preference, substrate affinity/capacity,

subcellular location, and activated conjugating moiety. ,

G)~Ll(U.\''OV\O~vll t.n:lI~JJJQ\G sJO')"tro,-ttS lncll...{(~,-{ - OH/ - COO-'~

~\-'\ ~ SIJ..\{c.\.A_-rut\'S\trOSG sujo\k,V-0c-l(!~ l'''C\C(W 9Nr'Q\

I

~r'\)4p~

Q) qhJ(UirtllnOS~I\ ft}Ll\,)~'R.(Ct)C/ ~\1\~,~iOn~ Ct'-C \'~15r-1H/ -"LDI'lU1l'\~I~l,"til -\:rr\ Y"\ Sulf!\'-tfCt 'lsfex(.tsc.S (JU~fCl t I'LlI'lJfqu ) l~ J')C,\v <:: l' a ff7 nll('L~

-V (GrXl C1-t:~) _ _ '~_ , '

(j) 0!l.\:cu'oV\05-~ --tIV!I'~)-f(i'a.s~s" 'f?u/\d II) t... E. ~\,)'\I(XO~O(f)'):)\)/ SU \ f-R -t. Y (N\\'.:>~"( Ctf c. S -f\.,,~ I\G\. \ () LGI \--t-O~Q \

@ G \"-'lLUY'()\,\O)<..,j \ -ty(.\ Yl stc rQ. \~, uct \~CA~~ 0 r{)O\ c-t'j .. U!'16 \~~Q.- 8\ ph~Jy.;~~'Y 'S\\.J lA) .. \r'()V; \\,. C,-( \ 0 ~ <o, 'J-o.. -ty' ()\'~fC:rt\fC,. o.c-ti~lo-te (A iy\0\ C~"-:J . ~ \ ?"D ?noC C\.(!\tfQ)it'Z 'J- \ plI=>SPhoSU\ fu--\.~. (PAP, ')

-------- 7

,

15

Nlarks

33. Some tll~ercul<:~sis patients (approx. 53%) respond to iso~a~id treatmentbetter than others. In these .pat1e~ts (i.e, those that respond better), plasma is.omazId concentrations measured 6 hours after ingesuon of a dose of 9.8 to 10.0 mg/ko body weight were found to be approx, 4.6 Jig/ml, while in the other 47% of the patients conc~ntrations averaged 1.1 pg/mJ. Explain why these differences occur.

5

\~Q'(\'\ O"C \c\ \ s 'SU.S LQP-tr bLL "*-D ~tCtko \'Ism b"-t N'D.~~ \ -tXQfLS\LrCtS-LS ~'-U- -txJ 1<:) CAml tLQ) .

~Q r-c:... CAI~ --tuJa 1~<'l!1X'P't1{P("~ 1 .~Olrl\..Q IrS O( w-alx-t,yl-tYCVIS-~rCQC ( )\AI) ti(\Ly~.eS: k~)C\11 Ul""\LA- ',JAI:L. V-lY!ILC . ,_ .,,-)\A;il_ i \ . ~y !OiC~"ULt " L~VI) -tC~-l CtlY\O~9J--t O\fkrL~ Vyll V\dltd \S./ \'-21'0) L Sho'-I\JS \/q)' 1 Ut.:?j)1

ulfl-ertifl-l t()8t ((1'\10rph\"SYYIJ d- t~ W¥1IL ~SL{I-t (,I)

.~ 010 SO\} C o. p\\.0 i'lO --tk/. r= s- ."" .

f- Ct )A-'-. 0._ LQ.~)J I D "-tot S .

\ n{PrfY'-Q dl Cl-lQ C~c{-b11 Gtov~

" ~ \ >() \.A..') .G.:cQ ty I c..{. :--'0(5,

-- \i~ c\ \f-Ki"C!/) UL \ s: q'L{Q \1 tu-UVG ((c. en itcv(J'\-t a{f,f) It..:J Y\ \J~ a ~ vtd Y\cyh bLf o-t til") 0Lj VN S') .

- ~ L-l1 /,. o-r e Let! e VI -t ~ Who hCl v c. low C(' Col') ce 11 t 1 Gi t ( ?'llJ

of Is()nl,C{Ll~ ec\'-tQy \0 h~lYS art lilt~ {'qs-t

Q U.tljl u."t.O :J'j ·tY\.Q. On.-LO J \ f"Y"'-et 0.100.1 \ c-e a V11o~

q,lA.-k~ Oc~ clQaVLd rf"b~I!M ~ -fbs-tQ ( -{1"'0ll"\ 1{)&trv1cktoLJ vJho oJl:'.- ~IDUJ~(V' QCQ -tA'J/CrW vS .

Isoniazid