Chapter 2
Design and Synthesis of Novel Spiroindoline Heterocycle Derivatives as Anti-Diabetic Agents
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
2.1. Introduction
Type 2 Diabetes is a metabolic disorder that is primarily characterized by insulin resistance, relative
insulin deficiency and hyperglycemia. It is very rapidly increasing in the developed world as well as in
India. The Western Pacific region and Europe have the highest number of people with diabetes,
approximately 67 and 53 million, respectively. The highest prevalence rates are found in North
America (9.2 percent) and Europe (8.4 percent). The five countries with the largest numbers of people
with diabetes are India, China, the United States, Russia and Germany. The fraction of Type 2
diabetics in other parts of the world varies substantially, almost certainly for environmental and
lifestyle reasons, though these are not known in detail. Diabetes affects over 150 million people
worldwide and this number is expected to double by 2025.1
Type-2 Diabetes is often associated with obesity, hypertension, elevated cholesterol (combined
hyperlipidemia), and with the condition often termed Metabolic syndrome. It is also associated with
acromegaly, Cushing's syndrome and a number of other endocrinological disorders. Additional factors
found to increase risk oftype-2 diabetes include aging2 and a less active lifestyle.3
The cornerstone of treatment of diabetes is life style modification through increased physical activity
and attention to food intake, particularly among the obese in which weight loss is the principal goal.
When life style modification do not result in normalization or near normalization of metabolic
abnormalities, pharmacologic therapy is required. Before 1995 sulfonylurea was the only anti-diabetic
agents available for the treatment oftype-2 diabetes. More than 50% of patients were treated with oral
monotherapy, 40% people were treated with insulin therapy and a small percentage taking
sulfonylurea along with insulin. After 1995 numbers of new classes of pharmacological agents were
introduced in the market. The classes currently available are insulin and insulin analogues for type-
1/type-2 diabetes sulfonylureas,4 glinides,5 biguinides,6 thiazolidinediones (glitazones)/ acarbose,
miglitol (a-glycosidase inhibitors)8 and metformin for type-2 diabetes (T2D).
The advent of the thiazolidinediones (TZD) anti-diabetic drugs (Rosiglitazone, Pioglitazone,
Ciglitazone, Troglitazone and Netoglitazone), has led to extensive research in the area of anti-diabetic
drug discovery and development. Later on, Glitazars, a new class of compounds showing anti
hyperglycemic activity, were developed. Glitazars are basically Glitazones with thiazoline-2,4-dione
ring replaced with acid derivatives (Figure 1 ). However, despite their effectiveness in treating
diabetes, these drugs possess undesirable side effects, such as increased adiposity, edema, significant
cardiac hypertrophy, and the risk of heart failure. Thus, there is a need to discover novel compound
with improved therapeutic profiles for the treatment of diabetes and related metabolic disorders.
60
Chapter2 ~ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
0
F ~NH ~ ~\(
0 \\ 0
Net~NH
cT JJ s-i 0 0
Ciglitazone
0
lL~NH N 0 \\
0
0~ "!~)(_" ~ \ 0 ~J-o \_d -N 0 0
Pioglitazone
~NH 0 0
HO Troglitazone
JTT 501
dL~OH "'N o~OP HN~- / --c
f' ~ ~ //
-0
Farglitazar
;s~o~oYooH o~ ,, I I
0 ~ 0 ~ 01
Tesaglitazar, AZ 242
Figure I. The antihyperglycemic g/itazones and glitazars
2.2. Basis of work
A careful survey of literature reveals that small molecules like phenyl acetic acid,9 cinnamic acid/0
hydrocinnamic acid,11 N-Benzoyl-DL-phenylalanine12 and l3-nitrostyrenes13 possess anti
hyperglycemic activity (Figure 2).
--- ... \
~r-CoH ~~~oo'H :V ,---- :v ,--,____ '-----
Figure 2. Small molecules as anti-hyperglycemic agents
\
~ I ~~ I
~COOH
V~,,'N~COPh
Within last few years many indole based carboxylic acid derivatives were reported as good anti-
diabetic agents. Most notably among these are 1-benzoyl-indole. carboxylic acid derivatives/4 2,3-
substituted indoyl-5yl acetic acid/5 oxazole based indole acetic acid derivative/6 pyrrolo[2,3-
b )pyridine based acids17 and indole-2-carboxamido-phenoxy acid derivatives. 18 A natural product
yohimbine19 have also been found to exhibit antidiabetic activity (Figure 3).
61
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009
Diabetic A ents
' ' I '
____;-7, COOH ,:
MoO~ '---·
)::oo Ph
,' :Hooc~ ' ... -_,
' I ' '
Figure 3. Indole based anti-hyperglycemic agents having acid functionality.
"::: .
H H . ·, MeOOC OH
Yohibine
A series of spirosuccinimide aldose reductase inhibitors were showing hypoglycaemic activity
comparable to that of ciglitazone (Figure 4).20
Figure 4. Anti-hyperglycemic spirosuccnimides
From figure 3, we concluded that all these anti-hyperglycemic compounds possess an indole with
acid functionality as the basic pharmacophore. Acid functionality is also present in small anti
hyperglycemic molecules (Figure 2). The concept of joining these two units a) the basic nucleus
indole and b) the side chain cinnamic acid, in a spiro framework came from spiro anti-hyperglycemic
agents. A careful examination of spirosuccinimide reveals that the spiro frameworks have been made
on phthalimide and saccharin. However, anti-hyperglycemic spiro frameworks built on indole have
not yet been reported. Thus, we have designed spiro [indoline-heterocycle] based carboxylic acid I
and its bioisostere nitro derivatives II as anti-diabetic agents (Figure 5). Our designed motif contains
62
Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
an indole based carboxylic acid (nitro) in a spiro system. Thus it contains the hybrid of all the
essential pharmacophoric features of the anti-hyperglycemic molecules.
Basic nudeus
UCOOH
R2
Side chain with acid functionality
c:/ -'-R2
::::,.._'
II
Spiro mode of arrangement
Figure 5. Designing of spiroindole based anti-hyperglycemic motif
2.3. Chemistry
c:/ -'-R2
::::,.._'
Bioisostere of acid
The retro-synthesis of the designed molecular scaffold suggests that it could be synthesized by a three
component coupling of isatin, proline and cinnamic acid or 13-nitrostyrene derivatives (Figure 6).
L-...-.-->
Figure 6. Retro-synthesis of designed molecular scaffold.
l>-cooH N H
Isatin derivatives were synthesized by Sandmeyer's method.21 Reacting the aniline 1 with chloral 2
and hydroxylamine hydrochloride 3 in the presence of Na2S04 to form compound 4, which was then
treated with PPA at 100 °C to yield isatin 5 (Scheme 1, Table 1 ).
R'O Na~04 R'O iNOH PPA RO=!O
I 0 + CCI3CHO + NH20H 10 NH 0 N 0
NH2 2 3 H 1 4 5
Scheme 1. Synthesis ofisatin derivatives
Table 1.
Entry R Compound Yield(%)
1 H Sa 65
2 Cl Sb 60
3 F Sc 70
4 N02 Sd 55
63
Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
Sandmeyer reaction is believed to proceed via following mechanism (Scheme 2).
OCxl NH CHO -2HCI
no ~ )l
NH CHO (±)
~ ail--t;J 0
4 H
H NH NH ~· HzO
~~~:A . I Q - t;J Q H H
Scheme 2. Mechanism of Sandmeyer isatin synthesis
~0
~N~o H 5
Substituted cinnamic acid esters were formed by esterification of cinnamic acid with alcohols in the
presence of catalytic amount of cone. H2S04 (Scheme 3, Table 2).
~COOH Cat. H2S0
4 ~COOR' -...;:::: + R10H
R ...-;:::.
7 R 6 8
Scheme 3. Synthesis of cinnamic acid esters
Table 2.
Entry R RI Compound Yield(%)
1 H Me Sa 70
2 H Et Sb 65
3 CF3 Me Sc 75
4 Cl Me Sd 75
5 MeO Me Se 68
6 MeO Et Sf 65
13-Nitrostyrenes were synthesized by reacting aromatic aldehydes with nitromethane in acetic acid.
The reaction was catalyzed by ammonium acetate and irradiated with ultrasound waves (Scheme 4,
Table 3).
RCHO
9 )))))
Scheme 4. Synthesis of substituted f3-nitrostyrene derivatives
64
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
Table 3.
Entry R Compound Yield(%)
4-F-CJI4 lOa 50
2 4-Me2NC6H4 lOb 40
3 3-MeOCJI4 tOe 40
4 2,5-Me02C6H3 lOd 45
5 4-H0-3-MeO C6H4 tOe 50
6 3,4-0CH20C6H3 lOf 60
Isatin derivative S was reacted with proline derivatives 11 and cinnamic acid esters S I ~-nitrostyrene
10 I cinnamaldehyde I cinnamic acid I a, ~-unsaturated ketones in aqueous ethanol to furnish
spiroindole derivatives 12 (Scheme 5, Table 4).
R2 :r /]
:::-... EWG t_)-cooH
~ ... ) R~O ~+ lh Et0H-H20 R1
+ N N 0 R1 H R
H 11 5 8 or 10 (X= CH2 11a, CHOH 11 b, S 11c)
Scheme 5. Synthesis if spiroindole derivatives 12
Table 4.
Entry Starting material Product Yield(%)
Isatin (R) Ester/Nitro/aldehyde/acid Proline (X)
(R1, R2, EWG)
1 Sa (H) Sa (H, H, COO Me) lla (CH2) 12a 80
2 Sa(H) Sb (H, H, COOEt) lla (CH2) 12b 78
3 Sa (H) Sc (4-CF3, H, COOMe) lla (CH2) 12c 95
4 Sa (H) Sd ( 4-Cl, H, COO Me) llc (S) 12d 90
5 Sa(H) Se (4-MeO, H, COOMe) llb (CHOH) 12e 65
6 Sa(H) Sb (H, H, COOEt) llb (CHOH) 12f 65
7 Sa (H) Sf(4-Me0, H, COOEt) lla (CH2) 12g 70
8 Sa (H) Sa (H, H, COO Me) llc (S) 12h 85
9 Sa(H) lOa (4-F, H, N02) lla (CH2) 12i 80
10 Sa(H) lOb (4-Me2N, H, N02) lla (CH2) 12j 60
Contn ....
65
Chapter2 ~ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
Continued from previous page ............
11 Sa (H) lOe (3-MeO, 4-HO, N02) lla (CH2) 12k 58
12 Sa (H) lOf (3,4-0CHzO, N02) lla (CHz) 121 55
13 Sa (H) lOd (2-MeO, 5-MeO, N02) lla (CH2) 12m 57
14 Sb (Cl) lOc (3-MeO, H, N02) lla (CH2) 12n 60
15 Sb (Cl) lOa (4-F, H, N02) lla (CH2) 12o 78
16 Sb (Cl) lOb (4-Me2N, H, NOz) lla (CH2) 12p 58
17 Sc (F) lOc (3-MeO, H, N02) lla (CH2) 12q 60
18 Sd (NOz)) lOc (3-MeO, H, N02) lla (CHz) 12r 55
19 Sa(H) lOc (3-MeO, H, N02) lla (CH2) 12s 60
20 Sa (H) (H,H,COOH) lla (CH2) 12t 15
21 Sa (H) (4-MeO, H, CHO) lla (CHz) 12u 30
22 Sa (H) (H, H, COCH3) lla (CHz) 12v 70
23 Sa (H) (H, H, COPh) lla (CHz) 12w 73
The three component coupling reaction of isatin, proline and olefm is believed to proceed through
1 ,3-dipolar cycloaddition by intermediacy of an azomethine ylid. Isatin reacts with proline to form an
intermediate 18 and/or 19. Intermediate 18 I 19 eliminates a molecule of C02 to generate another
intermediate azomethine ylid 20 which is then trapped by activated olefins (Scheme 6). The [3 + 2]
cycloaddition reaction occurs in a concerted manner thus stereochemistry of olefin is maintained in
the fmal product 12.
0 H•N--\ "'-':: HO t-1 U-1 u-
~ COOH ~HOOCb
0 h 0 N N H 11 H
5
f?J~ ~~v -C02
~N-b H .
18
0 HOOC ·~
b c{ ~_)
eel ~ """""I 0 h N 0
~ H
18 19
~~'R er=:=t<EWG
H 12
Scheme 6. Proposed mechanism for the synthesis of spiroindoles 12
66
Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
Compound 12 on treatment with a number of amino alkyl halides or acid halides in the presence of
K2C03 in dry DMF or acetone yields compound 13 (Scheme 7, Table 5).
12 +
13
Scheme 7.
Table 5.
Entry 12 R EWG 13 Yield(%)
12a eN\_ __
COO Me 13a 80
2 12b Cl~-- COOEt 13b 75
3 12b '~-N , COOEt 13c 77 I
4 12b ,~/'r- COOEt 13d 68
5 12b eN\_ __
COOEt 13e 80
6 12b eN\_ __
COOEt 13f 83
7 12b (Me2CHhNCH2CH2 --- COOEt 13g 70
8 12b 1\ COOEt 13h 74 0\.._/N\_ __
9 12b p-r\_/·, COOEt 13i 77
\__J
Cl 10 12b 4Me0-C6H4CO COOEt 13j 95
11 12g eN\_ __
COOEt 13k 69
Contn ......
67
Chapter 2 l::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
Continued from previous page .............
12 12h 1\ COO Me 131 75 0\__JN\_ __
13 12h eN\_ __
COO Me 13m 72
14 12s eN\_ __
N02 13n 75
15 12t eN\_ __
COOH 13o 76
16 12t ~ COOH 13p 75 _JN~--
We have performed the 1 ,3-dipolar cycloaddition reaction on 4-nitrobenzaldehyde. A mixture of 4-
nitrobenzaldehyde, proline and dimethyl maleate in ethanol was refluxed for 6 h. After usual work
up, two products were isolated and characterized as compound 15 and 16 in 3 : 1 ratio (Scheme 8).
Reflux O CHOC>-COOH
+ N 02N H
r(COOMe + ~ ----------
COOMe
Ethanol
15
SchemeS.
Formation of compound 15 and 16 in the reaction might be explained by the intermediacy of the
azomethine ylid 17 which is trapped by dimethyl maleate to form 15 and by 4-nitrobenzaldehyde to
form compound 16 (Scheme 9).
MeOOC'=/COOMe r 0}11 COOMe d OMe I h
~N ~N
15 17
Scheme 9.
osf' \-\ d'L
02N, "'-.7 . y 16 N02
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Chapter 2 Design and Synthesis of Novel Spiroindo/ine-Heterocyc/e Derivatives as Anti- 2009 Diabetic A ents
2.4. Experimental
2.4.1. Synthesis of Isatins (5)
Aniline (10 mmol), chloral hydrate (10 mmol) and hydroxylamine hydrochloride (10 mmol) were
taken in 30 ml aqueous sodium sulfate. The reaction mixture was refluxed for 2 h and then allowed to
cool down. The solid separated (isonitrosoacetanilide) filtered and dried. Isonitrosoacetanilide (5
mmol) and PPA (50 mmol) were taken in 100 ml round bottom flask and heated on a water bath for 6
h. The reaction was followed by TLC monitoring. After completion, the reaction mixture was poured
in ice water ( 1000 ml) with stirring. The solid separated was filtered off and crystallized from ethanol.
2.4.1.1. Characterization data for synthesized isatins
lsatin (Sa): Physical state: Orange red solid. Mp = 199 °C. ESI MS 148 (m/z) = (M + Ht. IR (KBr,
cm-1): 3446,3021, 1737, 1623, 1216.
5-Chloroisatin (Sb): Physical state: Orange red solid. Mp = 255 °C. ESI MS (m/z) = 181 (M + Ht. IR
(KBr, cm-1): 3621, 3020, 1746, 1700, 1615, 1216.
5-Fiuoroisatin (Sc): Physical state: Deep red solid. Mp = 225 °C. ESI MS (mlz) = 166 (M + Ht. IR
(KBr, cm.1): 3445, 3067, 1738, 1620, 1488, 1390.
5-Nitroisatin (Sd): Physical state: yellow solid. Mp = 248 °C. ESI MS (m/z) = 193 (M + Ht. IR
(KBr, cm.1): 3334,3095, 1772, 1752,1621,1531, 1469, 1338.
2.4.2. Synthesis of cinnamic acid esters (8)
Cinnamic acid (10 mmol) was taken in 30 ml of absolute ethanol (or methanol) and 2 drops of
concentrated sulphuric acid was added to it. Then it was refluxed over water bath for 2 h. The reaction
was followed by TLC monitoring. After completion, the reaction mixture was concentrated and 50 ml
water was added to it. It was then extracted with ethyl acetate. The ethyl acetate layer was washed by .
aqueous sodium hydrogen carbonate, then dried over anhydrous sodium sulphate and concentrated to
yield the cinnamic acid esters.
2.4.2.1. Characterization data for synthesized cinnamic acid esters
Methyl cinnamate (Sa): Physical state: oil. ESI MS (mlz) = 163 (M + Ht. IR (Neat, cm-1): 3062,
3029,2951,1719,1638,1578,1496,1449,1317.
Ethyl cinnamate (8b ): Physical state: oil. ESI MS (mlz) = 177 (M + Ht. IR (Neat, cm-1): 3042, 2972,
2856, 1719, 1631, 1573, 1444.
Methyl (4-trifluoromethyl) cinnamate (8c): Physical state: oil. ESI MS (m/z) = 245 (M + Ht. IR
(Neat, cm-1): 3045,2965,2829, 1725, 1639, 1618, 1485.
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Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
Methyl (4-chloromethyl) cinnamate (8d): Physical state: oil. ESI MS (m/z) = 197 (M + Ht. IR
(Neat, cm-1): 3029,2968,2827, 1724, 1640, 1605, 1480.
Methyl (4-methoxy) cinnamate (8e): Physical state: oil. ESI MS (mlz) = 193 (M + Ht. IR (Neat, cm-
1): 3022,2935,2856,1705,1634,1609,1508,1444.
Ethyl (4-methoxy) cinnamate (8t): Physical state: oil. ESI MS (m/z) = 207 (M + Ht. IR (Neat, cm-1):
2981,2840,1705,1634,1605,1513,1462.
2.4.3. Synthesis of ~-nitrostyrenes (10)
Aromatic aldehyde (10 mmol), nitromethane (50 mmol), ammonium acetate (10 mmol) and 10 ml of
acetic acid were taken in I 00 ml beaker. The reaction mixture was immerged in ultrasonic bath and
irradiated for 4-6 h. The reaction was followed by TLC monitoring. After completion, 50 ml of ice
water was added to it and the solid sepamted was filtered off and dried.
2.4.3.1. Characterization data for synthesized ~-nitrostyrenes
(E)-1-Fluoro-4-(2-nitrovinyl)benzene (lOa) Physical state: White solid. Mp = 134 °C. ESI MS (m/z)
= 168 (M + Ht. IR (KBr, cm-1): 3112,2946, 1639, 1610, 1494, 1428.
(E)-N,N-Dimethyl-4-(2-nitrovinyl)aniline (lOb): Physical state: White solid. Mp = 185 °C. ESI MS
(m/z) = 193 (M + Ht. IR (KBr, cm-1): 3106,2921, 1597, 1533, 1483, 1438, 1321.
(E)-1-Methoxy-3-(2-nitrovinyl)benzene (lOc): Physical state: White solid. Mp = 93 °C. ESI MS
(m/z) = 180 (M + Ht. IR (KBr, cm-1): 3105,2932, 1625, 1601.
(E)-1,4-Dimethoxy-2-(2-nitrovinyl)benzene (lOd): Physical state: White solid. Mp = 115 °C. ESI
MS (mlz) = 210 (M + Ht. IR (KBr, cm-1): 3106,2945, 1665, 1600.
(E)-2-Methoxy-4-(2-nitrovinyl)phenol (lOe): Physical state: White solid. Mp = 165 °C. ESI MS
(m/z) = 196 (M + Ht. IR (KBr, cm-1): 3473,3115,2970, 1672, 1603.
(E)-5-(2-Nitrovinyl)benzo[d][l,3]dioxole (lOt): Physical state: White solid. Mp = 153 °C. ESI MS
(m/z) = 193 (M + Ht. IR (KBr, cm-1): 3115, 2919, 1628, 1602, 1495, 1455.
2.4.4. Synthesis of spiro[indoline-heterocycle]-carboxylic acid I ester I aldehyde I ketones I nitro
derivatives (12)
Isatin (5 mmol), L-proline (7.5 mmol), cinnamic acid I esters/ cinnamaldehyde I a, ~-unsaturated
ketones I ~-nitrostyrenes (5 mmol) and 20 m1 aqueous ethanol (70%) were taken in 50 ml round
bottom flask. The reaction mixture was stirred for 4 h at room temperature. The reaction was
followed by monitoring. After completion, the solid separated was filtered off and dried. The filtrate
was concentrated and purified by silica-gel column chromatography. Compounds obtained from
filtration and from column were mixed and crystallized from methanol.
70
Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
2.4.5. Synthesis of spiro[indoline-heterocycle]-carboxylic acid derivatives (13)
In a 50 ml round bottom flask, compound 12 (5 mmol), alkyl halide (6 mmol), K2e03 (15 mmol) and
20 ml of dry DMF were taken. The reaction mixture was refluxed for 8-12 hover a water bath. The
reaction was followed by TLe monitoring. After completion, K2e03 was filtered off and 100 ml of
water was added to the filtrate. Then it was extracted with ethyl acetate. The ethyl acetate layer was
washed with water several times to remove DMF then dried over anhydrous sodium sulphate and
concentrated to yield crude. The crude was purified by basic alumina column chromatography.
2.4.6. Characterization data for synthesized spiro [indoline-heterocycle] derivatives
Methyl 2-oxo- 11-phenyl- 11,21,51,61,71,7a1-hexahydrospiro [indoline- 3,31-pyrrolizine]- 21-
carboxylate (12a)
Physical State: White solid. Mp = 195 °e. FAB MS (m/z) = 363 (M + Ht. IR (KBr, cm-1): 3158,3089,
2958, 2851, 1725, 1616, 1470, 1335, 1180, 746. 1H NMR (eDeh, 200 MHz) o = 1.68-2.03 (m, 4H),
2.60-2.70 (m, 2H), 3.11 (s, 3H), 3.66 (dd, J= 9.6 & 12.0 Hz, IH), 4.05-4.16 (m, 2H), 6.98-7.10 (m,
2H), 7.24-7.39 (m, 5H), 7.45-7.48 (m, 2H), 9.49 (s, 1H). Be NMR (eDel3, 50 MHz) o = 28.1, 31.7,
48.6, 51.8, 53.2, 62.3, 73.5, 74.3, 111.2, 122.4, 126.1, 126.5, 127.5, 128.3, 129.0, 130.0, 139.8, 142.1'
170.2, 182.0. Elemental analysis calculated for e22H22N20 3: e, 72.91; H, 6.12; N, 7.73. Found: e,
73.02; H, 6.10; N, 7.80.
Ethyl 2-oxo- 11 -phenyl-1 1,21 ,51 ,61, 71,7 a 1 -hexahydrospiro [indoline-3,3 1 -pyrrolizine ]-2 1 -carboxylate
(12b) Physical State: White solid. Mp = 160 °e. FAB MS (m/z) = 377 (M + Ht. IR (KBr, cm-1):
3160, 2965, 2871, 1724, 1614, 1470, 1341, 1176, 753. 1H NMR (eDeh, 200 MHz) o = 0.70 (t, J=
7.1 Hz, 3H), 1.65-2.02 (m, 4H), 2.58-2.70 (m, 2H), 3.52-3.71 (m, 3H), 4.10-4.16 (m, 2H), 7.01-7.09
(m, 2H), 7.21-7.38 (m, 5H), 7.46-7.49 (m, 2H), 10.23 (s, 1H). Be NMR (eDeh, 50 MHz) o = 13.7,
28.2, 31.8, 48.4, 53.2, 60.9, 62.0, 73.5, 74.3, 111.3, 122.4, 126.2, 126.6, 127.4, 128.3, 129.0, 130.0,
140.0, 142.4, 169.6, 182.1. Elemental analysis calculated for e23H2~203: e, 73.38; H, 6.43; N, 7.44.
Found: e, 73.24; H, 6.36; N, 7.32.
Methyl 2-oxo- 1 1-(4-(trifluoromethyl) phenyl)- 11,21,51 ,61, 71, 7a1-hexahydrospiro [indoline-3,31-
pyrrolizine]-21-carboxylate (12c) Physical State: White solid. Mp = 165 °e. FAB MS (m/z) = 431
(M + Ht. IR (KBr, cm-1): 3163, 2950, 1729, 1597, 1468, 1338, 1124, 745. 1H NMR (eDeh, 200
MHz) o = 1.63-2.04 (m, 4H), 2.60-2.71 (m, 2H), 3.13 (s, 3H), 3.74 (m, 1H), 4.08-4.15 (m, 2H), 7.00-
7.11 (m, 2H), 7.26-7.33 (m, 2H), 7.46-7.50 (m, 2H), 7.65-7.74 (m, 2H), 9.79 (s, IH). Be NMR
(eDel3, 50 MHz) o = 25.6, 29.9, 46.8, 50.3, 51.3, 60.7, 71.7, 72.6, 109.7, 120.9, 123.4, 124.7, 127.9,
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Chapter2 ~ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
128.5, 129.9, 130.1 139.3, 140.4, 168.3, 180.1. Elemental analysis calculated for C22H19F3N20 3: C,
63.46; H, 4.60; N, 6.73. Found: C, 63.31; H, 4.72; N 6.58.
Methyl 7'-(4-chlorophenyl)- 2-oxo- 3' ,6', 7', 7a'-tetrahydro-1 'H-spiro [indoline-3,5'-pyrrolo [1,2-
c]thiazole]-6'-carboxylate (12d) Physical State: White solid. Mp > 225 °C. ESI MS (m/z) = 415 (M +
Ht. IR (KBr, cm"1): 3415, 3203, 3096, 2921,2883, 1723, 1616, 1487, 1440, 1328, 1231, 1192, 1135,
1089, 804, 738, 700. 1H NMR (Pyridin-d5, 300 MHz) () = 2.30-2.37 (m, 2H), 3.05 (d, J = 10.2 Hz,
IH), 3.30 (d,J= 10.2 Hz, IH), 3.38-3.42 (m, IH), 3.69-3.70 (m, IH), 3.78 (d,J= 12.2 Hz, IH), 4.61
(s, 3H), 6.34-6.37 (m, 2H), 6.77-6.90 (m, 4H), 7.18 (t, J = 7.1 Hz, 2H), 11.33 (s, IH). Elemental
analysis calculated for C21 H19CIN20 3S: C, 60.79; H, 4.62; N, 6.75. Found: C, 60.82; H, 4.51; N, 6.65.
Methyl 6'-hydroxy- 1 '-(4-methoxyphenyl)-2-oxo- 1 ',2' ,5' ,6', 7' ,7a'-hexahydrospiro [indoline-3,3'
pyrrolizine]-2'-carboxylate (12e) Physical State: White solid. Mp > 225 °C. FAB MS (m/z) = 409
(M + Ht. IR (KBr, cm-1): 3516,3194,3032,2917,2837, 1740, 1694, 1616, 1513, 1470, 1174,756.
1H NMR (CDCh, 200 MHz) () = 2.12-2.20 (m, 2H), 2.59-2.80 (m, 2H), 3.23 (s, 3H), 3.58-3.62 (m,
IH), 3.80 (s, 3H), 3.99-4.12 (m, 2H), 4.49-4.51 (m, lH), 6.95-7.11 (m, 2H), 7.23-7.49 (m, 7H), 9.98
(s, lH). 13C NMR (CDCh, 50 MHz) cS = 56.4, 56.9, 57.3, 60.1, 60.4, 60.7, 66.8, 76.2, 76.5, 115.3,
119.3, 126.4, 130.6, 131.1, 132.5, 133.9, 134.7, 136.5, 137.0, 147.9, 163.7, 174.9, 184.5. Elemental
analysis calculated for C23H24N20 5: C, 67.63; H, 5.92; N, 6.86. Found: C, 67.58; H, 6.00; N, 6.70.
Ethyl 6'-hydroxy-2-oxo-1'-phenyl-1',2',5',6',7',7a'-hexahydrospiro[indoline-3,3'-pyrrolizine]-2'
carboxylate (12t) Physical State: White solid. Mp > 225 °C. FAB MS (m/z) = 393 (M + Ht. IR
(KBr, cm-1): 3505, 3172, 2968, 2866, 1725, 1612, 1469, 1353, 1170. 1H NMR (CDC13, 200 MHz) () =
0.70 (t, J= 7.1 Hz, 3H), 2.17-2.22 (m, 2H), 2.58-2.84 (m, 2H), 3.54-3.64 (m, 3H), 4.00-4.15 (m, 2H),
4.49-4.52 (m, lH), 6.96-7.09 (m, 2H), 7.22-7.45 (m, 7H), 8.96 (s, lH). 13C NMR (CDC13, 50 MHz) cS
= 14.7, 45.8, 50.3, 54.2, 61.9, 64.6, 66.8, 73.5, 74.4, 110.7, 120.4, 124.2, 125.5, 127.2, 127.9, 128.0,
132.2, 141.1, 143.0, 169.9, 179.1. Elemental analysis calculated for C23H2~204: C, 70.39; H, 6.16;
N, 7.14. Found: C, 70.28; H, 6.06; N, 7.00.
Ethyl 1 '-( 4-methoxyphenyl)-2-oxo-1 ',2' ,5' ,6', 7', 7 a' -hexahydrospiro [indoline-3,3 '-pyrrolizine ]-2 '
carboxylate (12g) Physical State: White solid. Mp = 205 °C. ESI MS (m/z) = 407 (M + Ht. IR (KBr,
cm-1): 3227, 2963, 2867, 1738, 1615, 1515, 1469, 1383, 1249, 1176, 1022, 758. 1H NMR (CDCh,
200 MHz) cS = 0.70 (t, J= 7.1 Hz, 3H), 1.64-2.01 (m, 4H), 2.59-2.68 (m, 2H), 3.51-3.70 (m, 3H), 3.79
(s, 3H), 4.04-4.15 (m, 2H), 6.91 (d, J = 8.6 Hz, 2H), 6.99-7.09 (m, 2H), 7.23-7.41 (m, 4H), 9.82 (s,
lH). 13C NMR (CDCh, 50 MHz) cS = 13.7, 28.1, 31.7, 48.4, 52.5, 55.6, 60.8, 62.2, 73.4, 74.2, 111.1,
72
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
114.5, 122.4, 126.3, 126.6, 129.3, 130.0, 131.9, 142.3, 159.0, 169.7, 181.8. Elemental analysis
calculated for C24H26Nz04: C, 70.92; H, 6.45; N, 6.89. Found: C, 70.84; H, 6.35; N, 7.00.
Methyl 2-oxo-71-phenyl-31 ,61 ,71, 7a 1-tetrahydro-1 1H-spiro(indoline-3,51-pyrrolo(1,2-c]thiazole)-61
-
carboxylate (12h) Physical State: White solid. Mp = 230 °C. ESI MS (mlz) = 381 (M + Ht. IR (KBr,
cm-1): 3438, 3249, 3030, 2945, 2856, 1744, 1708, 1615, 1470, 1438, 1385, 1332, 1180, 1131, 756,
722, 702. 1H NMR (Pyridin-d5, 300 MHz) () = 2.19- 2.34 (m, 2H), 3.00 (d, J = 10.2 Hz, lH), 3.19-
3.26 (m, 2H), 3.59-3.62 (m, lH), 3.72 (d, J = 12.2 Hz, lH), 4.37 (s, 3H), 6.29-6.35 (m, 2H), 6.52-6.65
(m, 4H), 6.82- 6.89 (m, 2H), 7.14 (d, J = 7.3 Hz, lH), 11.23 (s, lH). 13C NMR (CDC13, 50 MHz) 8 =
36.3, 51.1, 51.6, 54.4, 59.7, 73.7, 75.5, 110.1, 121.1, 123.7, 127.5, 127.6, 128.2, 129.1, 130.4, 139.4,
143.0, 169.3, 178.4. Elemental analysis calculated for C21H20N20 3S: C, 66.29; H, 5.30; N 7.36. Found:
C, 66.22; H, 5.41; N, 7.24.
1 1-(4-Fiuorophenyl)-21-nitro-1 1,21 ,51,61 ,71, 7a1-hexahydrospiro(indoline-3,31-pyrrolizin]-2-one (12i)
Physical State: White solid. Mp > 225 °C. ESI MS (m/z): 368 (M + Ht. IR (KBr, cm-1): 3430, 3021,
2975, 1619, 1549, 1514, 1216,761,670. 1H NMR (DMSO-d6, 300 MHz) 8 = 1.36-1.66 (m, 2H), 1.89-
2.01 (m, 2H), 2.50-2.61 (m, 2H), 4.55-4.64 (m, 2H), 6.40 (t, J = 10.0 Hz, lH), 6.67 (d, J = 7.4 Hz,
1H), 6.96-7.02 (m, 3H), 7.18-7.23 (m, 3H), 7.87 (d, J= 7.1 Hz, 1H), 10.31 (s, lH). Elemental analysis
calculated for C20H18FN30 3: C, 65.39; H, 4.94; N, 11.44, Found: C, 65.42; H, 5.00; N, 11.30.
1 1-( 4-(Dimethylamino )phenyl)-2 1-nitro-1 1,21,51 ,61, 71
, 7a 1-hexahydrospiro(indoline-3,3 1-pyrrolizin]-
2-one (12j) Physical State: White solid. Mp = 204 °C. ESI MS (m/z) = 393 (M + Ht. IR (KBr, cm-1):
3185, 3021, 2943,2832, 1616, 1546, 1470, 1327, 1215, 762, 700. 1H NMR (DMSO-~, 300 MHz) 8 =
1.36-1.64 (m, 2H), 1.90-1.99 (m, 2H), 2.50-2.58 (m, 2H), 2.76 (s, 6H), 4.40 (d, J = 10.9 Hz, 1H), 4.55-
4.60 (m, lH), 6.21-6.27 (m, 1H), 6.48 (d,J= 8.1 Hz, 2H), 6.67 (d,J= 8.9 Hz, 1H), 6.93-7.03 (m, 3H,
ArH), 7.17-7.21 (m, lH), 7.78 (d, J= 7.2 Hz, 1H), 10.34 (s, 1H). Elemental analysis calculated for
C22H24N40 3: C, 67.33; H, 6.16; N, 14.28. Found: C, 67.28; H, 6.20; N, 14.35.
1'-(4-Hydroxy-3-methoxyphenyl)-21-nitro-1', 21, 51
, 61, 7', 7a1-hexahydrospiro(indoline-3, 3 1
-
pyrrolizin]-2-one (12k) Physical State: White solid. Mp > 225 °C. ESI MS (mlz) = 396 (M + Ht. IR
(KBr, cm-1): 3511,3318,3030,2968,2832, 1621, 1550, 1509, 1478. 1H NMR (DMSO-d6, 300 MHz)
() = 1.37-1.66 (m, 2H), 1.90-2.03 (m, 2H), 2.50-2.63 (m, 2H), 3.49 {s, 3H), 4.44 (d, J = 10.7 Hz, 1H),
4.60-4.64 (m, 1H), 6.34 (dd,J= 10.0 & 10.7 Hz, 1H), 6.57-6.67 (m, 4H), 7.05 (t,J= 7.2 Hz, 1H), 7.23
(t, J= 7.3 Hz, 1H), 7.83 {d, J = 7.1 Hz, lH), 8.90 (s, 1H), 10.24 (s, 1H). Elemental analysis calculated
for C21H21N30 5: C, 63.79; H, 5.35; N, 10.63. Found: C, 63.65; H, 5.30; N, 10.50.
11 -(Benzo(d] (1,3]dioxol-5-yl)-21-nitro-1' ,21 ,51 ,61, 71
, 7a 1-hexahydrospiro(indoline-3,31-pyrrolizin]-2-
one (121) Physical State: White solid. Mp > 225 °C. FAB MS (m/z) = 394 (M + Ht. IR (KBr, cm-1):
73
Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
3186, 3084, 1704, 1621, 1550, 1500, 1473, 1254, 1041, 751. 1H NMR (CDCh + DMSO-d6, 200 MHz)
8 = 1.41-1.47 (m, lH), 1.70-1.76 (m, lH), 1.90-2.09 (m, 2H), 2.55-2.77 (m, 2H), 4.42 (d, J= 10.8 Hz,
lH), 4.68-4.72 (m, lH), 5.88 (s, 2H), 6.16-6.26 (m, lH), 6.59 (s, 2H), 6.67-6.71 (m, 2H), 7.00 (t, J =
7.5 Hz, lH), 7.23 (t, J = 7.4 Hz, lH), 7.66 (d, J = 7.4 Hz, lH), 10.24 (s, lH). 13C NMR (CDCh +
DMSO-d6, 50 MHz) 8 = 30.7, 33.0, 56.2, 57.1, 68.6, 75.3, 79.9, 97.1, 106.3, 113.5, 115.3, 126.9,
127.4, 130.3, 131.6, 135.0, 135.1, 148.5, 152.3, 152.7, 182.9. Elemental analysis calculated for
C21H19N30 5: C 64.12; H, 4.87; N, 10.68. Found: C, 64.20; H, 4.72; N, 10.50.
1 '-(2,5-Dimethoxyphenyl)-2'-nitro-1 ',2' ,5' ,6', 7', 7a '-hexahydrospiro(indoline-3,3'-pyrrolizin ]-2-
one (12m) Physical State: White solid. Mp >225 °C. FAB MS (m/z) = 410 (M + Ht. IR (KBr, cm-1):
3192, 3082, 2969, 1705, 1618, 1541, 1501, 1471, 1254, 1212, 1049, 745. 1H NMR (CDC13 + DMSO
~. 200 MHz) 8 = 1.41-1.47 (m, 2H), 1.95-2.05 (m, 2H), 2.55-2.78 (m, 2H), 3.51(s, 3H), 3.66 (s, 3H),
4.70-4.75 (m, 1H), 5.24 (d, J = 10.8 Hz, lH), 6.23 (t, J = 10.4 Hz, 1H), 6.65-6.63 (m, 3H), 7.00-6.93
(m, 2H), 7.18 (t, J= 7.6 Hz, lH), 7.59 (d, J= 7.5 Hz, 1H), 10.24 (s, 1H). 13C NMR (CDCh + DMSO
~. 75 MHz) 8 = 17.7, 24.2, 26.7, 37.8, 42.0, 49.8, 54.2, 54.9, 55.2, 62.4, 73.5, 91.4, 108.5, 111.4,
111.8, 113.2, 120.0, 121.6, 123.5, 126.6, 128.8, 141.9, 150.6, 151.9, 176.5. Elemental analysis
calculated for C22H23N30s: C, 64.54; H, 5.66; N, 10.26. Found: C, 64.42; H, 5.70; N 10.20.
5-Chloro-1 '-(3-methoxyphenyl)-2'-nitro-1 ', 2', 5', 6', 7', 7a'-hexahydrospiro(indoline-3,3'
pyrrolizin]-2-one (12n) Physical State: White solid. Mp > 225 °C. ESI MS (m/z) = 414 (M + Ht. IR
(KBr, cm-1): 3447, 2970, 2838, 1616, 1587, 1546, 1480, 1449, 1370, 1269, 1207, 1049, 822, 774. 1H
NMR (DMSO-d6, 300 MHz) 8 = 1.38-1.64 (m, 2H), 1.90-1.97 (m, 2H), 2.50-2.62 (m, 2H), 3.59 (s,
3H), 4.57-4.61 (m, 2H), 6.30-6.36 (m, 1H), 6.63- 6.77 (m, 4H), 7.10 (s, 1H), 7.26 (d, J = 6.7 Hz, 1H),
8.07 (s, lH), 10.41 (s, lH). 13C NMR (DMSO-d6, 75 MHz) 5 = 25.6, 28.1, 51.0, 51.1, 55.1, 63.5, 74.9,
90.8, 111.4, 113.3, 114.5, 120.5, 126.1, 127.3, 127.7, 129.9, 130.1, 135.1, 142.4, 159.3, 177.4.
Elemental analysis calculated for C21H20ClN30 4: C, 60.95; H, 4.87; N, 10.15. Found: C,, 60.86; H,
4.78; N, 10.28.
5-Chloro-1 '-( 4-fluorophenyl)-2'-nitro-1 ',2' ,5' ,6', 7', 7a '-hexahydrospiro[indoline-3,3 '-pyrrolizin]-
2-one (12o) Physical State: White solid. Mp > 225 °C. ESI MS (mlz) = 402 (M + Ht. IR (KBr, cm-1):
3399, 3029, 2979, 2822, 1620, 1545, 1515, 1470, 1233, 1195, 844, 676. 1H NMR (DMSO-~, 300
MHz) 8 = 1.37-1.66 (m, 2H), 1.92- 1.98 (m, 2H), 2.50-2.61 (m, 2H), 4.58-4.68 {m, 2H), 6.31-6.38 (m,
1H), 6.67 (d, J = 7.9 Hz, 1H), 7.06 (d, J = 7.8 Hz, 2H), 7.24 (s, 3H), 8.08 {s, 1H), 10.43 (s, 1H).
Elemental analysis calculated for C2oH17ClFN30 3: C, 59.78; H, 4.26; N, 10.46. Found: C, 59.69; H,
4.18; N, 10.55.
74
Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
5-Chloro-1'-(4-(dimethylamino)phenyl)-2'-nitro-1',2',5',6',7',7a'-hexahydrospiro[indoline-3,3'
pyrrolizin]-2-one (12p) Physical State: White solid. Mp = 210 °C. ESI MS (mlz) = 427 (M + Ht. IR
(KBr, cm-1): 3429, 3078, 1616, 1550, 1530, 1478, 1444, 1368, 1331, 1199, 822. 1H NMR (DMSO-d6,
300 MHz) o = 1.31-1.61 (m, 2H), 1.70-1.95 (m, 2H), 2.46-2.50 (m, 2H), 2.78 (s, 6H), 4.45-4.57 (m,
2H), 6.20-6.26 (m, lH), 6.52 (d, J = 7.2 Hz, 2H), 6.66 (d, J = 7.7 Hz, 1H), 6.98 (d, J = 7.4 Hz, 2H),
7.26 (d, J = 7.0 Hz, lH), 8.02 (s, lH), 10.39 (s, 1H). Be NMR (DMSO-d6, 75 MHz) o = 25.6, 27.9,
50.9, 50.9, 63.4, 74.8, 91.4, 111.3, 112.5, 120.0, 126.0, 127.5, 127.7, 129.1, 130.0, 142.5, 150.1,
177.7. Elemental analysis calculated for C22H23ClN40 3: C, 61.90; H, 5.43; N, 13.12. Found: C, 68.59;
H, 6.88; N, 11.30.
5-Fiuoro-1'-(3-methoxyphenyl)-2'-nitro-1', 2', 5', 6', 7', 7a'-hexahydrospiro[indoline-3, 3'
pyrrolizin]-2-one (12q) Physical State: White solid. Mp > 225 °C. ESI MS (m/z) = 398 (M + Ht. IR
(KBr, cm-1): 3431, 3021, 2975, 1605, 1546, 1486, 1215, 1043, 761, 671. 1H NMR (DMSO-~, 300
MHz) o = 1.35-1.64 (m, 2H), 1.92-2.00 (m, 2H), 2.46-2.50 (m, 2H), 3.59 (s, 3H), 4.56-4.59 (m, 2H),
6.38 (t, J = 10.1 Hz, lH), 6.63-6.78 (m, 4H), 7.05-7.10 (m, 2H), 7.93 (d, J = 7.7 Hz, lH), 10.32 (s,
lH). Be NMR (DMSO-~, 75 MHz) o = 25.5, 28.1, 50.9, 51.1, 55.2, 63.5, 75.1, 91.0, 110.8, 113.2,
114.6, 115.4, 116.5, 120.7, 129.9, 135.2, 139.7, 159.4, 177.6. Elemental analysis calculated for
C21 H20FN30 4: C, 63.47; H, 5.07; N, 10.57. Found: C, 63.50; H, 4.98; N, 10.40.
1'-(3-Methoxyphenyl)-2',5-dinitro-1',2',5',6',7',7a'-hexahydrospiro[indoline-3,3'-pyrrolizin]-2-
one (12r) Physical State: White solid. Mp > 225 °C. ESI MS (m/z) = 425 (M + Ht. IR (KBr, cm-1):
3418, 3020, 2975, 1605, 1525, 1451, 1333, 1216, 1106, 1045, 762, 671. 1H NMR (DMSO-~, 300
MHz) o = 133-1.67 (m, 2H), 1.90-2.02 (m, 2H), 2.50-2.63 (m, 2H), 3.56 (s, 3H), 4.55-4.64 (m, 1H),
4.83 (d, J= 10.0 Hz, 1H), 6.41 (dd, J= 10.0.& 10.6 Hz, lH), 6.70-6.78 (m, 3H), 6.87 (d, J= 8.7 Hz,
1H), 7.11 (t,J= 7.7 Hz, lH), 8.19 (dd,J= 2.0 & 8.7 Hz, lH), 8.94 (d, J= 2.0 Hz, 1H), 11.02 (s, 1H).
Elemental analysis calculated for C21H2oN40 6: C, 59.43; H, 4.75; N, 13.20. Found c. 59.52; H, 4.70;
N, 13.32.
1 '-(3-Methoxyphenyl)-2'-nitro-1' ,2' ,5' ,6', 7' ,7a'-hexahydrospiro[indoline-3,3'-pyrrolizin]-2-one
(12s) Physical State: White solid. Mp > 225 °C. ESI MS (m/z) = 380 (M + Ht. IR (KBr, cm-1): 3454,
2966, 2848, 1615, 1585, 1532, 1482, 1464. 1H NMR (DMSO-~, 300 MHz) o = 1.38-1.64 (m, 2H),
1.90-1.97 (m, 2H), 2.50-2.62 (m, 2H), 3.59 (s, 3H), 4.57-4.61 (m, 2H), 4.92 (d, J= 13.1 Hz, 1H), 6.75-
6.82 (m, 2H), 7.10-7.15 (m, 2H), 7.26 (d, J= 6.9 Hz, lH), 7.37-7.45 (m, 3H), 10.50 (s, !H). Elemental
analysis calculated for C21H21N304: C, 66.48; H, 5.58; N, 11.08. Found: C, 66.56; H, 5.47; N, 11.98.
75
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
2-0xo-1 '-phenyl-1 ',2 ',5' ,6', 7', 7a '-hexahydrospiro[indoline-3,3 '-pyrrolizine]-2'-carboxylic acid
(12t) Physical State: White solid. Mp = 225 °e. FAB MS (mlz) = 349 (M + Ht. IR (KBr, cm-1): 3460,
3130, 3077, 2969, 1738, 1614, 1472, 1370, 752. 1H NMR (DMSO-d6, 300 MHz) o = 1.61-1.82 (m,
4H), 2.35-2.45 (m, 2H), 3.59 (dd, J = 9.6 & 12.3 Hz, 1H), 3.66-3.73 (m, 1H), 3.80 (d, J = 12.3 Hz,
1H), 6.84 (d, J= 7.5 Hz, lH), 7.00 (t, J= 7.5 Hz, 1H), 7.25 (t, J= 7.5 Hz, 2H), 7.31-7.38 (m, 5H),
10.35 (s, 1H), 12.00 (s, lH). Be NMR (DMSO-d6, 75 MHz) o = 27.6, 30.6, 47.6, 52.2, 61.2, 72.7,
72.9, 110.1, 121.31, 126.1, 126.8, 127.0, 127.9, 128.8, 129.6, 140.7, 143.1, 170.8, 179.4. Elemental
analysis calculated for C21 H20N20 3: e, 72.40; H, 5.79; N 8.04. Found: e, 72.30; H, 5.72; N 7.90.
1'-(4-Methoxyphenyl)-2-oxo-1',2',5',6',7',7a'-hexahydrospiro[indoline-3,3'-pyrrolizine]-2'-
carbaldehyde (12u) Physical State: White solid. Mp = 188 °C. FAB MS (m/z) = 363 (M + Ht. IR
(KBr, cm-1): 3224,2961, 1721, 1616, 1468, 1246,753. 1H NMR (CDCh, 200 MHz) o = 1.72-1.96 (m,
4H), 2.58-2.66 (m, 2H), 3.01 (d, J= 13.6 Hz, 1H), 3.88 (s, 3H), 4.07-4.12 (m, lH), 4.78-4.82 (m, 1H),
6.75-7.19 (m, 4H), 7.24-7.42 (m, 4H), 9.12 (s, lH), 9.54 (s, 1H). Be NMR (eDeh + DMSO-~, 75
MHz) o = 28.6, 31.8, 45.0, 48.5, 55.8, 67.2, 72.1, 73.5, 111.2, 111.7, 121.4, 122.7, 124.2, 125.6, 126.8,
128.5, 129.2, 130.1, 141.8, 157.9, 181.3, 199.3. Elemental analysis calculated for C22H22N20 3: C,
72.91; H, 6.12; N 7.73. Found: e, 72.80; H, 5.98; N 7.56.
2'-Acetyl- 1'-phenyl- 1',2',5',6',7',7a'-hexahydrospiro [indoline- 3,3'-pyrrolizin] -2-one (12v)
Physical State: white solid. Mp = 225 °e. FAB MS (m/z) = 347 (M + Ht. IR (KBr, cm-1): 3050,2954,
2846, 1720, 1608, 1511, 1430. 1H NMR (eDeh, 300 MHz) o = 1.64 (s, 3H), 1.80-2.03 (m, 4H), 2.55-
2.69 (m, 2H), 3.65-3.72 (m, 1H), 4.07-4.15 (m, 1H), 4.27 (d, J= 11.9 Hz, 1H), 7.02-7.11 (m, 2H),
7.22-7.46 (m, 7H), 9.74 (s, 1H). Be NMR (eDeh, 75 MHz) o = 26.1, 28.6, 29.7, 46.6, 51.2, 68.2,
71.4, 72.0, 109.7, 121.3, 121.3, 124.1, 125.6, 125.7, 126.6, 127.4, 128.5, 138.5, 139.8, 180.3, 202.5.
Elemental analysis calculated for C22H22N20 2: C, 76.28; H, 6.40; N, 8.09. Found: C, 76.20; H, 6.28; N
7.94.
2'-Benzoyl-1 '-( 4-methoxyphenyl)-1 ',2' ,5' ,6', 7' ,7a '"-hexahydrospiro[indoline-3,3'-pyrrolizin]-2-one
(12w) Physical State: white solid. Mp = 210 °e. FAB MS (m/z) = 439 (M + Ht. IR (KBr, cm-1): 3038,
2930, 2846, 1715, 1610, 1522, 1442. 1H NMR (CDCh, 300 MHz) o = 1.71-2.06 (m, 4H), 2.59-2.72
(m, 2H), 3.75 (s, 3H), 3.85-3.92 (m, lH), 4.18-4.26 (m, 1H), 4.91 (d, J = 11.4 Hz, 1H), 6.61 (d, J =
7.6Hz, 1H), 6.87 (d, J= 8.6Hz, 2H), 7.04 (t, J= 7.1 Hz, 1H), 7.09-7.17 (m, 3H), 7.25-7.33 (m, 2H),
7.38-7.46 (m, 4H), 8.72 (s, 1H). Be NMR (CDeh, 75 MHz) o = 25.9, 29.3, 46.9, 50.9, 53.9, 63.2,
70.6, 72.4, 108.9, 112.8, 120.9, 123.8, 126.2, 126.5, 126.7, 127.07, 128.0, 130.4, 131.5, 135.8, 139.5,
76
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
157.2, 180.0, 195.5. Elemental analysis calculated for C28H26N20 3: C, 76.69;, H, 5.98; N, 6.39. Found:
C, 76.50; H, 6.06; N 6.28.
Methyl 2-oxo-1 '-phenyl-1-(2-(piperidin-1-yl)ethyl)-1 ',2' ,5' ,6', 7', 7a '-hexahydrospiro(indoline-3,3 '
pyrrolizine)-2'-carboxylate (13a) Physical State: Oil. ESI MS (m/z) = 474 (M + Ht. IR (Neat, cm-1):
2950, 2869, 1724, 1608, 1464, 1371, 1323, 1192, 756. 1H NMR (CDCh, 200 MHz) 8 = 1.44-1.61 (m,
8H), 1.86-2.04 (m, 2H), 2.55-2.63 (m, 10H), 3.05 (s, 3H), 3.58-3.63 (m, lH), 4.05-4.11 (m, 2H), 6.95
( d, J = 7.9 Hz, lH), 7.02-7.09 (m, 1 H), 7.23-7.36 (m, 5H), 7.42-7.46 (m, 2H). 13C NMR (CDCb, 50
MHz) 8 = 24.6, 26.2, 28.0, 30.1, 31.0, 31.5, 38.4, 48.5, 51.6, 53.3, 55.2, 55.7, 62.6, 73.3, 108.9, 122.3,
125.9, 126.5, 127.5, 128.4, 128.7, 129.06, 140.1, 144.0, 159.1, 170.1, 178.9. Elemental analysis
calculated for C29H35N303: C, 73.54; H, 7.45; N, 8.87. Found: C, 73.50; H, 7.55; N 8.78.
Ethyl 1-(2-chloroethyl)- 2-oxo- 1 '-phenyl- 1 ',2',5',6',7',7a'-hexahydrospiro (indoline- 3,3'
pyrrolizine)-2'-carboxylate (13b) Physical State: Oil. FAB MS (m/z) = 439 (M + Ht. IR (Neat, em· 1): 3040, 2970, 2856, 1725, 1613, 1465, 1351, 1178. 1H NMR (CDCh, 200 MHz) 8 = 0.69 (t, J= 7.1
Hz, 3H), 1.65-2.02 (m, 4H), 2.27-2.30 (m, 2H), 2.38-2.70 (m, 4H), 3.50-3.72 (m, 3H), 4.08-4.15 (m,
2H), 7.03- 7.10 (m, 2H), 7.21-7.39 (m, 5H), 7.45-7.50 (m, 2H). 13C NMR (CDCb, 50 MHz) 8 = 13.7,
25.9, 29.2, 40.9, 43.1, 51.0, 52.9, 53.2, 60.6, 61.7, 76.4, 110.4, 121.8, 123.3, 126.9, 128.6, 129.5,
132.0, 136.8, 144.6, 161.3, 168.6, 174.0. Elemental analysis calculated for C25H27ClN20 3: C, 68.41;
H, 6.20; N, 6.38. Found: C, 68.31; H, 6.30; N, 6.28.
Ethyl 1-(2-( dimethylamino) ethyl)-2-oxo- 1 '-phenyl- 1 ',2' ,5' ,6', 7', 7a '-hexahydrospiro (indoline-
3,3'-pyrrolizine)-2'-carboxylate (13c) Physical State: Oil. FAB MS (m/z) = 448 (M +H)+. IR (Neat,
cm-1): 3070, 2960, 2852, 1725, 1614, 1472, 1346. 1H NMR (CDCb, 200 MHz) 8 = 0.70 (t, J = 7.1
Hz, 3H), 1.85-2.02 (m, 4H), 2.25 (s, 6H), 2.45- 2.55 (m, 6H), 3.44-3.54 {m, 3H), 3.97-4.03 (m, 2H),
6.83 (d, J = 7.8 Hz, 1H), 6.95-6.97 (m, lH), 7.15-7.28 {m, 5H), 7.35-7.39 (m, 2H). Elemental
analysis calculated for C21H33N30 3: C, 72.46; H, 7.43; N, 9.39. Found: C,,72.39; H, 7.35; N, 9.26.
Ethyl 1-(1-( dimethylamino) propan-2-yl)- 2-oxo-1 '-phenyl- 1 ',2' ,5' ,6', 7', 7a '-hexahydrospiro
[indoline-3,3 '-pyrrolizine )-2 '-carboxylate (13d)
Physical State: Oil. FAB MS (m/z) = 462 (M + Ht. IR (Neat, cm-1): 3078, 2970, 2846, 1724, 1614,
1475, 1348. 1H NMR (CDCh, 200 MHz) 8 = 0.65 {t, J = 7.1 Hz, 3H), 1.65-2.10 (m, 4H), 2.18 (d, J =
3.8 Hz, 3H), 2.26 (s, 3H), 2.29 (s, 3H), 2.42-2.48 (m, 4H), 2.99-3.10 (m, lH), 3.46-3.57 (m, 3H), 3.98-
4.04 {m, 2H), 6.97-7.01 (m, 2H), 7.20-7.30 (m, 5H), 7.36-7.40 {m, 2H). 13C NMR (CDCh, 50 MHz) 8
= 11.6, 13.7, 28.2, 31.7, 41.1, 41.2, 43.9, 48.3, 53.5, 56.9, 60.6, 62.2, 73.3, 73.5, 109.0, 122.4, 126.2,
77
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
126.5, 127.4, 128.4, 129.0, 130.0, 140.2, 144.4, 169.8, 179.1. Elemental analysis calculated for
C28H35N30 3: C, 72.86; H, 7.64; N, 9.10. Found: C, 72.80; H, 7.50; N, 8.98.
Ethyl 2-oxo-1 1 -phenyl- 1-(2-(pyrrolidin-1-yl) ethyl)- 11,21,51,61, 71, 7a 1- hexahydrospiro [indoline-
3,31-pyrrolizine]-21-carboxylate (13e)
Physical State: Oil. FAB MS (m/z) = 474 (M + H/. IR (Neat, cm-1): 3050, 2955, 2868, 1725, 1615,
1469, 1340, 1174. 1H NMR (CDC13, 200 MHz) o = 0.57 (t, J= 7.1 Hz, 3H), 1.70-2.02 (m, 8H), 2.48-
2.59 (m, 6H), 2.76 (t, J= 7.6 Hz, 2H), 3.44-3.54 (m, 3H), 3.97-4.03 (m, 2H), 6.88 (d, J = 7.8 Hz, lH),
6.97-7.01 (m, lH), 7.14-7.28 (m, 5H), 7.35-7.39 (m, 2H). Elemental analysis calculated for
C29H3sN303: C, 73.54; H, 7.45; N, 8.87. Found: C, 73.51; H, 7.36; N, 8.78.
Ethyl 2-oxo-1 1-phenyl-1-(2-(piperidin-1-yl) ethyl)-1 1,21,51 ,61, 71, 7a 1-hexahydrospiro [indoline-
3,31-pyrrolizine]-21-carboxylate (13f) Physical State: Oil. FAB MS (m/z) = 488 (M + H/. IR (Neat,
cm-1): 3051, 2955, 2866, 1725, 1613, 1472, 1349. 1H NMR (CDC13, 200 MHz) 8 = 0.65 (t, J = 7.1
Hz, 3H), 1.26-1.59 (m, 8H), 1.64-1.88 (m, 2H), 2.49-2.64 (m, lOH), 3.51-3.58 (m, 3H), 4.05-4.10 (m,
2H), 6.93 (d, J= 7.5 Hz, lH), 7.05-7.09 (m, 1H), 7.23-7.44 (m, 5H), 7.45-7.52 (m, 2H). 13C NMR
(CDCh, 50 MHz) o = 13.8, 26.6, 26.3, 28.0, 31.3, 31.5, 38.5, 48.4, 53.4, 55.1, 55.9, 60.6, 62.1, 73.1,
73.3, 108.9, 122.3, 126.1, 126.5, 127.4, 128.4, 129.0, 129.9, 140.1, 144.2, 159.0, 169.8, 178.8.
Elemental analysis calculated for C3oH37N30 3: C, 73.89; H, 7.65; N, 8.62. Found: C, 73.80; H, 7.56;
N, 8.50.
Ethyl 1-(2-( diisopropylamino )ethyl)-2-oxo- 1 1-phenyl-1 1,21,51 ,61, 71, 7a 1-hexahydrospiro [indoline-
3,31-pyrrolizine]-21-carboxylate (13g) Physical State: Oil. FAB MS (m/z) = 504 (M + H/. IR (Neat,
cm-1): 2969, 2872, 1724, 1609, 1486, 1466, 1364, 1215, 1170, 757. 1H NMR (CDCh, 200 MHz) o =
0.67 (t, J= 7.1 Hz, 3H), 1.08 (d, J= 6.2 Hz, 12H), 1.62-1.92 (m, 4H), 2.57-2.72 (m, 4H), 3.04-3.10
(m, 2H), 3.49-3.60 (m, 3H), 3.75-3.79 (m, 2H), 4.05-4.11 (m, 2H), 6.90 (d, J= 7.9 Hz, IH), 7.04-7.08
(m, 1H), 7.23-7.33 (m, 5H), 7.43-7.47 (m, 2H). Elemental analy~is calculated for C31H41N30 3: C,
73.92; H, 8.20; N, 8.34. Found: C, 73.80; H, 8.26; N, 8.48.
Ethyl 1-(2-morpholinoethyl)-2-oxo- 1 1-phenyl-1 ',2' ,5' ,6', 7', 7a 1-hexahydrospiro [indoline-3,3 '
pyrrolizine]-21-carboxylate (13h) Physical State: Oil. FAB MS (m/z) = 490 (M + H/. IR (Neat, cm-
1): 3048, 2856 1724, 1613, 1522, 1470, 1346, 1178. 1H NMR (CDCh, 200 MHz) 0 = 0.57 (t, J =
7.1Hz, 3H), 1.78-1.95 (m, 4H), 2.43-2.61 (m, IOH), 3.43-3.63 (m, 7H), 3.97-4.02 (m, 2H), 6.90 (d, J
= 7.7 Hz, IH), 7.05-7.07 (m, lH), 7.23-7.46 (m, 5H), 8.00- 8.03 (m, 2H). 13C NMR (CDC13, 50 MHz)
8 = 13.8, 28.0, 31.6, 38.0, 48.4, 53.4, 54.1, 55.5, 60.5, 62.3, 67.4, 73.2, 73.3, 108_8, 122.4, 126.1,
78
Chapter 2 l:_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
126.6, 127.7, 128.4, 129.0, 129.9, 140.1, 144.0, 169.7, 178.8. Elemental analysis calculated for
C29H3sN304: C, 71.14; H, 7.21; N, 8.58. Found: C, 71.20; H, 7.14; N 8.46.
Ethyl 1-(3-( 4-(3-chlorophenyl) piperazin-1-yl) propyl)-2-oxo- 1 '-phenyl- 1.' ,2 ',5' ,6', 7', 7a '
hexahydrospiro [indoline-3,3'-pyrrolizine)-2'-carboxylate (13i) Physical State: Oil. ESI MS (mlz)
= 613 (M + Ht. IR (Neat, cm.1): 2993,2876,2823, 1715, 1598, 1566, 1492, 1376, 1221, 1102, 1030,
947,858, 756. 1H NMR (CDCh, 200 MHz) 8 = 0.65 (t, J= 7.1 Hz, 3H), 2.04-1.86 (m, 6H), 2.59-2.45
(m, 10H), 3.20-3.18 (m, 4H), 3.63-3.49 (m, 3H), 4.13-4.02 (m, 2H), 6.95-6.77 (m, 4H), 7.37-7.02 (m,
7H), 7.47-7.43 (m, 2H). 13C NMR (CDCh, 50 MHz) 8 = 13.9, 25.2, 28.1, 31.6, 38.7, 48.4, 49.1, 53.4,
53.8, 55.7, 60.6, 62.2, 73.2, 130.5, 73.4, 108.9, 114.2, 116.1, 119.6, 122.3, 126.1, 126.6, 127.4, 128.4,
129.0, 129.9, 135.3, 140.1, 144.3, 152.7, 169.7, 178.9. Elemental analysis calculated for
C36~1ClN403 : C, 70.51; H, 6.74; N, 9.14. Found: C, 70.48; H, 6.60; N, 9.04.
Ethyl 1-( 4-methoxybenzoyl)- 2-oxo-1 '-phenyl- 1' ,2' ,5' ,6', 7', 7a '-hexahydrospiro [indoline-3,3 '
pyrrolizine)-2'-carboxylate (13j) Physical State: White solid. Mp = 143 °C. FAB MS (m/z) = 511
(M + Ht. IR (Neat, cm.1): 2968,2867, 1769, 1740, 1671, 1603, 1510, 1464, 1337, 1262, 1160, 1027,
841,762. 1H NMR (CDCb, 200 MHz) 8 = 0.74 (t, J= 7.1 Hz, 3H), 1.66-2.01 (m, 4H), 2.67-2.70 (m,
2H), 3.52-3.66 (m, 3H), 3.89 (s, 3H), 4.04-4.16 (m, 2H), 6.95-6.99 (d, J= 8.7 Hz, 2H), 7.18-7.44 (m,
8H), 7.76-7.89 (m, 3H). Elemental analysis calculated for C31H3oN20s: C, 72.92; H, 5.92; N, 5.49.
Found: C, 72.86; H, 6.00; N, 5.40.
Ethyl 1 '-(4-methoxyphenyl)-2-oxo- 1-(2-(piperidin-1-yl) ethyl)-1 ',2' ,5' ,6', 7', 7a '-hexahydrospiro
[indoline-3,3'-pyrrolizine)-2'-carboxylate (13k) Physical State: Oil. FAB MS (m/z) = 518 (M +
Ht. IR (Neat, cm-1): 2940, 2860, 1714, 1611, 1514, 1486, 1444, 1332, 1275, 1178, 1128, 755. 1H
NMR (CDCh, 200 MHz) 8 = 0.65 (t, J = 7.1 Hz, 3H), 1.20-1.60 (m, 8H), 1.81-1.91 (m, 2H), 2.45-
2.65 (m, lOH), 3.46-3.58 (m, 3H), 3.79 (s, 3H), 4.02-4.04 (m, 2H), 6.86-6.93 (m, 3H), 7.05-7.08 (m,
lH), 7.27-7.39 (m, 4H). 13C NMR (CDCh, 50 MHz) 8 = 13.8, 24.6, 27.9, 30.0, 31.3, 31.5, 38.4, 48.4,
52.7, 55.1, 55.6, 55.8, 60.6, 62.2,-73.2, 108.9, 114.4, 122.3, 126.5, 129.3, 129.9, 132.0, 144.1, 159.0,
169.8, 178.8. Elemental analysis calculated for C31H39N304: C, 71.93; H, 7.59; N, 8.12. Found: C,
71.86; H, 7.50; N, 8.19.
Methyl 1-(2-morpholinoethyl)-2-oxo-7'-phenyl-3' ,6', 7', 7a '-tetrahydro-1 'H-spiro[indoline-3,5'
pyrrolo[1,2-c)thiazole)-6'-carboxylate (131) Physical State: White solid. Mp = 95 °C. ESI MS (mlz)
= 494 (M + Ht. IR (KBr, cm-1): 3030, 2945, 2856, 1744, 1708, 1615, 1470, 1438, 1385, 1332, 1180,
1131, 756, 722, 702. 1H NMR (Pyridin-d5, 300 MHz) 8 = 2.19-2.34 (m, 2H), 2.43-2.61 (m, 8H), 3.19-
3.26 (m, 2H), 3.37 (s, 3H), 3.43-3.63 (m, 5H), 3.97-4.02 (m, 2H), 6.29- 6.35 (m, 2H), 6.52-6.65 (m,
79
Chapter2 ~ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
4H), 6.82-6.89 (m, 2H), 7.14 (d, J = 7.3 Hz, lH). 13C NMR (CDCh, 50 MHz) 8 = 28.0, 36.3, 38.3,
51.1, 51.6, 53.1, 54.4, 59.7, 73.7, 75.5, 110.1, 121.1, 123.7, 127.5, 127.6, 128.2, 129.1, 130.4, 139.4,
143.0, 169.3, 178.4. Elemental analysis calculated for C27H31N30 4S: C, 65.70; H, 6.33; N, 8.51.
Found: C, 65.62; H, 6.41; N, 8.42.
Methyl 2-oxo-7'-phenyl-1-(2-(pyrrolidin-1-yl)ethyl)-3 ',6', 7', 7a '-tetrahydro-1 'H-spiro[indoline-
3,5'-pyrrolo[1,2-c]thiazole]-6'-carboxylate (13m) Physical State: White solid. Mp = 90 °C. ESI MS
(m/z) = 478 (M + Ht. IR (KBr, cm-1): 3030,2927,2856, 1730, 1608, 1463, 1361, 1182, 1031,756. 1H
NMR (CDCh 300 MHz) 8 = 1.60-1.80 (m, 4H), 2.52-2.99 (m, 12H), 3.51 (s, 3H), 3.68-3.72 (m, lH),
3.85-3.96 (m, 2H), 6.92 (d,J= 7.8 Hz, lH), 6.99-7.04 (m, lH), 7.27-7.38 (m, 4H), 7.48 (d,J= 7.3 Hz,
2H), 7.68 (d, J= 7.5 Hz, lH). 13C NMR (CDCh, 75 MHz) 8 = 22.1, 22.3, 28.3, 35.3, 38.3, 49.9, 50.4,
51.0, 53.0, 53.9, 58.8, 72.4, 74.2, 106.9, 121.6, 122.2, 126.2, 126.9, 127.5, 128.8, 137.3, 142.1, 167.9,
176.3. Elemental analysis calculated for C27H31N30 3S: C, 67.90; H, 6.54; N, 8.80. Found: C, 67.82; H,
6.50; N, 8.70.
1 '-(3-Methoxyphenyl)- 2'-nitro-1- (2-(piperidin-1-yl) ethyl)-1 ',2 ',5' ,6', 7', 7a '- hexahydrospiro
[indoline-3,3'-pyrrolizin]-2-one (13n) Physical State: White solid. Mp = 160 °C. ESI MS (m/z) = 491
(M + Ht. IR (KBr, cm-1): 3030,2940, 1606, 1546, 1492, 1464, 1371, 1181, 1115, 1036, 747, 694: 1H
NMR (DMS0-<4, 300 MHz) 8 = 1.31-1.66 (m, 6H), 1.72-1.96 (m, 4H), 2.15-2.50 (m, 6H), 3.25-3.42
(m, 4H), 3.51 (s, 3H), 4.43 (d, J= 10.2 Hz, lH), 4.70-4.73 (m, lH), 6.22-6.25 (m, lH), 6.47 (s, lH),
6.61 (d, J = 7.8 Hz, 2H), 6.74 (d, J = 7.9 Hz, lH), 6.94-6.99 (m, lH), 7.09 (t, J = 7.4 Hz, lH), 7.22-
7.25 (m, lH), 7.66-7.68 (m, lH). 13C NMR (DMSO-~, 75 MHz) 8 = 22.6, 25.6, 25.8, 27.9, 37.6, 51.0,
52.8, 54.5, 54.9, 55.3, 64.1, 74.4, 124.7, 91.0, 109.1, 113.5, 113.7, 120.1, 122.3, 126.4, 129.4, 130.0,
134.3, 144.2, 159.3, 175.6. Elemental analysis calculated for C28H3~404: C, 68.55; H, 6.99; N, 11.42.
Found: C, 68.59; H, 6.88; N, 11.30.
2-0xo-1 '-phenyl-1-(2-(piperidin-1-yl) ethyl)-1 ',2' ,5,',6', 7', 7a '- hexahydrospiro [indoline-3,3'
pyrrolizine]-2'-carboxylic- acid (l3o) Physical State: White solid. Mp = 132 °C. FAB MS (m/z) = 460
(M + Ht. IR (KBr, cm-1): 3364, 2860, 1720, 1618, 1468, 1337, 1165, 748. 1H NMR (CDCh, 200
MHz) 8 = 1.45-1.32 (m, 6H), 1.85-2.04 (m, 4H), 2.14-2.27 (m, 6H), 2.62-2.65 (m, 4H), 3.59-3.68 (m,
3H), 4.07-4.13 (m, 2H), 6.95-7.08 (m, 2H), 7.23-7.38 (m, 5H), 7.43-7.47 (m, 2H), 9.78 (s, 1H).13C
NMR (CDCh, 50 MHz) 8 = 23.9, 25.6, 27.6, 31.1, 48.1, 52.9, 54.4, 56.0, 61.7, 61.8, 73.0, 73.7, 110.9,
121.9, 125.8, 126.9, 127.0, 127.9, 128.6, 129.7, 139.5, 142.2, 169.3, 181.3. Elemental analysis
calculated for C28H33N30 3: C, 73.18; H, 7.24; N, 9.14. Found: C, 73.12; H, 7.30; N, 9.22.
80
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
. 1-(2-(Diethylamino )ethyl)-2-oxo- 1 1-phenyl-1 1,21,51 ,61, 71
, 7a 1- hexahydrospiro [indoline-3,3 1-
pyrrolizine]-21-carboxylic acid (15p) Physical State: White solid. Mp = 125 °C. FAB MS (m/z) =
448 (M + Ht. IR (KBr, cm-1): 3410, 3067, 2968, 1719, 1612, 1468, 1337, 1169, 750. 1H NMR
(eDel3, 200 MHz) o = 0.90 (t, J= 7.1 Hz, 6H), 1.68-2.13 (m, 6H), 2.40 (q, J= 7.1 Hz, 4H), 2.63-
2.70 (m, 2H), 3.58-3.67 (m, 3H), 4.08-4.15 (m, 2H), 7.00-7.10 (m, 2H), 7.24-7.38 (m, 5H), 7.44-7.48
(m, 2H), 10.10 (bs, 1H). Be NMR (eDeh, 50 MHz) o = 12.0, 28.0, 31.6, 47.7, 48.5, 50.2, 53.3, 62.1,
62.7, 73.3, 74.1, 111.2, 122.4, 126.2, 126.6, 127.4, 128.3, 129.0, 130.1, 139.9, 142.5, 169.7, 181.7,
Elemental analysis calculated for e 27H33N30 3: e, 72.46; H, 7.43; N, 9.39. Found: e, 72.48; H, 7.50;
N,9.24.
Dimethyl3-(4-nitrophenyl)hexahydro-1H-pyrrolizine-1,2-dicarboxylate (15) Physical State: Oil.
FAB MS (m/z) = 349 (M + Ht. IR (KBr, cm-1): 3079, 2931, 2875, 1736, 1604, 1524, 1445, 1348,
1288, 856, 757. 1H NMR (eDeh, 200 MHz) o = 1.98-2.19 (m, 4H), 2.58-2.71 (m, 2H), 3.08 (s, 3H),
3.24-3.37 (m, 2H), 3.59 (d, J= 9.2Hz, lH), 3.73 (s, 3H), 4.03-3.97 (m, 1H), 7.50 (d, J= 7.8 Hz, 2H),
8.19 (d,J= 7.8 Hz, 2H). Be NMR (eDeh, 75 MHz) o = 22.5, 23.0, 42.2, 43.2, 48.9, 50.1, 58.2, 63.2,
71.5, 122.0, 127.1, 146.0, 149.6, 169.2, 171.3. Elemental analysis calculated fore 17H20N20 6: e, 58.61;
H, 5.79; N, 8.04. Found: e, 58.48; H, 5.70; N, 7.92.
2,3-Bis(4-nitrophenyl)hexahydropyrrolo[2,1-b]oxazole (16) Physical State: Oil. FAB MS (m/z) =
356 (M + Ht. IR (KBr, cm-1): 3080, 2943, 2862, 1601, 1520, 1347, 1203, 1086, 832.747, 695. 1H
NMR (eDeh, 200 MHz) o = 1.90- 2.25 (m, 4H), 2.83-2.88 (m, 1H), 3.14-3.20 (m, 1H), 3.87 (d, J =
7.9 Hz, 1H), 4.72 (d, J= 7.9 Hz, lH), 5.33-5.36- (m, 1H), 7.36-7.43 (m, 4H), 8.17-8.23 (m, 4H). Be
NMR (eDeh, 75 MHz) o = 24.4, 31.9, 52.6, 56.2, 78.8, 87.8, 99.7, 124.2, 124.3, 127.4, 128.4, 130.1,
145.9, 147.9, 148.3, 148.5, 166.0. Elemental analysis calculated for e 18H17N30 5: e, 60.84; H, 4.82; N,
11.83. Found: e, 60.72; H, 4.74; N, 11.72.
81
Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
2.5. Biological activity
2.5.1. Anti-diabetic activity
2.5.1.1. Animals: Adult male and female albino rats (Sprague Dawley) of body weight 160±20g,
bred in CDRI animal house were used during the course of experiment; 6 animals were kept in one
cage. All the animals were fed ad-lib standard pellet diet (Lipton, Bombay) and allowed unrestricted
access to water. The following norms were followed for animal room environment. Temperature: 22
± 1 ·c; Humidity: 50-50%; Light 300 Lux at floor level with regular 12 hours light cycle; noise level
50 decibels; ventilation 10-50 air changes per hour.
The blood-glucose lowering effects of the test samples/standard drugs were examined in the following
experimental models.
2.5.1.2. Sucrose--loaded rat model (SLM): Overnight fasted male Sprague Dawley rats were used for
the sucrose-loaded experiment. Blood was collected at 'O'min from the tail vein of the animals. After
the '0' min blood collection, samples/drugs were given to the test group consisting of 5 rats by oral
gavage at a dose of 100 mg/kg. Half an hour post test sample treatment, a sucrose-load of 10.0 gm/kg
body weight was given to each rat. The blood was collected at 30, 60, 90 & 120 min post sucrose-load.
2.5.1.3. Sucrose-challenged Streptozotocin-induced Diabetic rat Model (STZ); Male albino rats of
Sprague Dawley strain of body weight 160 ± 20 g were selected for this study. Streptozotocinl3
(Sigma, USA) was dissolved in 100 mM citrate buffer pH 4.5 and calculated amount of the fresh
solution was injected to overnight fasted rats (45 mglkg) intraperitoneally. Blood glucose level was
checked 48 h later by glucostrips and animals showing blood glucose values between 144 to 270 mg/dl
(8 to 15 mM) were included in the experiment and termed diabetic. The diabetic animals were divided
into groups consisting of five to six animals in each group. Animals of experimental groups were
administered suspension of the desired test samples orally (made in 1.0% gum acacia) at a dose of 100
mg/kg body weight. Animals of control gi:oup were given an equal amount of 1.0% gum acacia. A
sucrose load of2.5glkg body w~ight was given after 30 minutes of compound administration. After 30
minutes of post sucrose load blood glucose level was again checked by glucostrips at 30, 60, 90, 120,
180, 240, 300 min and at 24 h, respectively. Animals not found diabetic after 24 hours post treatment
of the test sample were not considered and omitted from the calculations and termed as non
responders. The animals, which did not show any fall in blood glucose profile in a group while the
others in that group, showed fall in blood glucose profile were also considered as non-responders.
Food but not water was withheld from the cages during the experimentation. Comparing the AUC of
82
LJzabetzc Agents I
experimental and control groups determined the percent anti-hyperglycemic activity. Statistical
comparison between groups was made by Student's 't' test.
%Anti-hyperglycemic activity= 100 -(AlB) x 100
A = A varage blood glucose level of test substance treated group at test time
B = A varage blood glucose level of control group at test time
2.5.1.4. Anti-hyperglycemic Activity in db/db mice: The db/db mouse is a well-characterized model
of type 2 diabetes. The background for the db/db mouse is the C57BL/Ks strain. The major deficiency
of the C57BL/KsBom-db mouse (db/db) is lack of a functionalleptin receptor. This leads to defective
leptin signalling and a complete lack of feedback from leptin. Both hypothalamic NPY content and
secretion are consequently elevated, and this result in hyperphagia and decreased energy expenditure,
obesity, insulin-resistance, hyperinsulinemia, hyperglycaemia and dyslipidemia. The db/db mouse
develops NIDDM from around week 10. The disease is stable until week 20, where destruction of
pancreatic 13-cells can be recognized clinically as decreasing levels of plasma insulin and very severe
hyperglycaemia. The db/db mouse has a maximal life span of9 -12 months. The male mice are more
diabetic than, and will normally die earlier than the females. The advantage of using male mice for
experimental purposes is that the fluctuations in plasma parameters are less than in the females where
the oestrogen cycle affects the clinical diabetes. The optimal age of db/db mice used for experiments
will be from week 12 to 18 when they have developed NIDDM with diabetic dyslipidemia but still
have functionalJ3-cells in the pancreas. C57BL/KsBom-db mice 12- 18 weeks, 40- 50 g bred in the
animal house ofCDRI, Lucknow. 10 mice (5 males and 5 females) were used in the experiments. The
mice were housed in groups of 5 (same sex) in a room controlled for temperature (23 ± 2.0 oq and
12/12 hours light/dark cycle (lights on at 6.00 am). Body weight was measured daily from day 1 to day
10. All animals had free access to fresh water and to normal chow except on the days of the
postprandial protocol day 6 and d~g the overnight fast before the OGTT on day 10. The animals
always had access to water during experimental periods. Blood glucose was checked every morning
up till day 5. On day 6 postprandial protocol was employed, in this method blood glucose was checked
at --0.5h and Oh Test drugs were given to the treatment group whereas vehicle received only gum
acacia (1.0%); the blood glucose was again checked at 1, 2, 3, 4 and 6h post test drug treatment. On
day 8, blood was collected for serum insulin measurements and fmally on day 10 an oral glucose
tolerance test (OGTT) was performed after an overnight fasting. Blood glucose was measured at -30.0
min and test drugs were administered. The blood glucose was again measured at 0.0 min post
treatment and at this juncture glucose solution was given at a dose of 3 gm!kg to all the groups
including vehicle. The blood glucose levels were checked at 30 min, 60 min, 90 min and 120 min post
83
Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
glucose administration. Quantitative glucose tolerance of each animal was calculated by area under
curve (AUC) method (Prism softwere). Comparing the AUC of experimental and control groups
determined the% anti-hyperglycemic activity. Statistical comparison was made by Dunnett's test.
2.5.1.5. Lipid estimation
2.5.1.5.1. Cholesterol: Cholesterol was estimated by the kit provided Roche Diagnostics. Cholesterol
esters are enzymatically hydrolyzed by cholesterol esterase (CE) to cholesterol and free fatty acids.
Free cholesterol, including the originally present, is then oxidized by cholesterol oxidase (CO) to
cholest-4-en-3-one and hydrogen peroxide. The hydrogen peroxide combines with hydroxyl benzoic
acid (HBA) and 4-amonoantipyrene in the presence of peroxidase (POD) to form a chromophore
(quinoneimine dye), which may be quantitated at 500-505 nm. The intensity of red colour formed is
directly proportional to the total cholesterol in the specimen and measured spectrophotometrically: 22
2.5.1.5.2. Triglycerides: Triglicerides were estimated using the kit provided by Reche Diagnostics.
Lipoprotein hydrolyses triglicerides to yield glycerol and fatty acids. Glycerol kinase converts glycerol
to glycerol -3-phosphate, which is oxidized by glycerol phosphate oxidase to dihydroxy acetone
phosphate and hydrogen peroxide. In the presence of peroxidase, hydrogen peroxide oxidatively
couples with 4-aminoantipyrene and 4-chlorophenol to produce red quinonimine dye. The intensity of
the red colour formed is directly proportional to concentration of triglycerides in the specimen and is
measured photometrically.
2.5.1.5.3. HDL-Cholesterol: HDL-Cholesterol was estimated using the kit provided by Roche
Diagnostics. Cholestest N HDL is a liquid reagent that directly measures the HDL -cholesterol
concentration in serum by new method that is based on the selective solubilising effect of proprietarY
detergent to the different lipoproteins. In the assay system, only HDL is solubilised by a special
detergent; other lipoproteins are not disrupted. After HDL is selectively disrupted, HDL cholesterol is
measured enzymatically.Z3
84
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
Table 6: In vivo anti-hyperglycemic activity profile of active compounds in SLM and STZ-S models
Example Anti-hyperglycemic activity (In vivo)
SLM STZ-S Model
5 hr 24hr ED so (mglk:g)
12a 15.8 ND ND
12b 15.9 ND ND
12c 45.5 28.8 32.1 62.8
12d 17.6 ND ND
12e 16.3 ND ND
12f 12.5 ND ND
12g 21.9 ND ND
12h 27.9 15.9 17.1
12i 15.8 ND ND
12j 35.9 22.8 23.2
12k 21.3 ND ND
121 21.9 ND ND
12m 54.5 32.8 37.1 39.8
12n 12.5 ND ND
12o 16.3 ND ND
12p 27.9 15.9 17.1
12q 17.6 ND ND
12r 16.9 ND ND
12s 26.8 13.5 13.5
12t 28.8 22.5 20.5
12u 10.0 ND ND
12v 10.5 ND ND
12w 6.8 ND ND
13a 43.5 20.7 23.8
13b 16.9 ND ND
13c 26.8 13.5 13.5
13d 19.2 ND ND
13e 23.1 ND ND
85
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
Continued from previous page .........
13f 35.2 14.1 16.5
13g 38.9 12.5 10.6
13h 20.5 ND ND
13i 38.1 22.0 24.1
13j 39.0 18.1 20.5
13k 31.4 17.5 18.0
131 38.9 16.1 18.4
13m 22.5 12.4 14.0
13o 54.0 25.9 30.0 89.9
13p 36.7 . 12.4 13.9
Table 7: In vivo anti-hyperglycemic, lipid lowering and triglyceride lowering profile of
active compounds 12c, 12m & 13o in db/db mice models(% efficacy, 100 mglkg)
Compound Anti-hyperglycemic Lipid lowering activity
activity (% Change) (%Change)
Day6 Day 10 TG CHOL. HDL
12c 32.8 44.5 -31.2 -24.4 +13.6
12m 38.0 43.7 -25.2 -17.8 +9.6
13o 37.0 45.8 -24.5 -20.6 +11.0
Dose dependant anti-hyperglycemic effect of 12c, 12m & 13o on sucrose challenged
streptozotasin-induced diabetic rats
Dose response cutve of 12c, 12m & 13o were obtained by administering different doses of test
compound to streptozotasin-induced diabetic rats (Table 8). Doses ranges from 7.5-100 mg!Kg were
given and blood glucose level was measured at 30, 60, 90, 120, 180, 240, 300, 1440 min post
administration of sucrose load as described in sucrose challenged streptozotasin-induced diabetic rats.
Compound 12c, 12m & 13o showed dose dependency and their ED50 were found to be 62.8 mg/Kg,
39.6 mg/Kg, & 89.9 mg!Kg respectively.
86
Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
Table 8. Dose dependent anti -hyperglycemic effect of 12c, 12m & 13o
Compound Dose (mg/kg) % glucose lowering
12c 7.5 13 .7
15 21.7*
25 26 .6**
50 32.8**
100 36.8***
12m 7.5 12.5
15 18.5
25 22.5*
50 28.5**
100 32.8**
13o 7.5 12.7
15 17 .0
25 21.7*
50 27 .6**
100 31.9**
Values are expressed as mean ± SD, N=5, p<0.05 (*), p<O.Ol (**) & p<0.001 (***) vs control.
Statistical analysis was made by Dunnett test (Prism Software).
35
30
i' 25
.5. 5l 20 0 u ::l
Cl 15 "0 0 0 iii 10
5
-20 0
Dose dependent anti-hyperglycemic effect of 12c
30 60 90 120 180 240 300 1440
Time (min)
••
-.- 12c 100mg/Kg
--12c 50mg/Kg
12c 25mg/Kg
-- 12c 15mg/Kg
-- 12c 7.5mg/Kg
---- Control
87
Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
35
~ 30
.§.. 25 Q)
~ 20 0
:I 15 C)
-g 10 ..2 tel 5
Dose dependent anti-hyperglycemic activity of 12m
- 12m 100 mg/Kg
1- 12m 50 mg/kg
12m 25 mg/Kg
-- 12m 15 mg/Kg
- 12m 7.5 mg/Kg
- contro l
0 ~----------------------------------------
-20 0 30 60 90 120 180 240 300 1440
Time (min)
Dose dependent anti-hyperglycemic effect of 13o
35
30
i" 25 .§.
--13o 100mg/Kg
5l 20 8 :::J
0, 15 "C 0 0 iii 10
5
0 L_ ________________________________________________ _
-20 0 30 60 90 120 180 240 300 1440
Time (min)
--- 13o 50mg/Kg
13o 25mg/Kg
---- 13o 15mg/Kg
-- 13o 7.5mg/Kg
---Control
88
Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
2.6. Conclusion
We have designed spiro [indole-heterocycle] as a common core motif based on the structures of
known anti-hyperglycemic agents. The designed motif is hybrid of indole and cinnamic acid
derivatives. These two subunits were arranged in a spiro system. The designed compounds were
synthesized via a three component coupling reaction of isatin derivatives, proline derivatives and
electron deficient alkenes like cinnamic acid esters, P-nitrostyrenes, cinnamaldehyde and chalcones.
We synthesized a series of spiro [indolene-heterocycle]-carboxylic acid, ester, carboxaldehyde,
carbonyl and nitro derivatives. The compounds were evaluated for their anti-hyperglycemic activity
in sucrose loaded model (SLM), sucrose challenged streptozotasin induced diabetic rat model (STZ)
and db/db mice model. Spiro [indolene-heterocycle]-carboxylic acid, ester and nitro derivatives were
showing significant blood glucose lowering activity in SLM model however, corresponding aldehyde
and ketones 12u, 12v and 12w were inactive. Compounds showing promising blood glucose lowering
activity in SLM model were further screened for their anti-hyperglycemic activity in STZ model.
Three compounds methyl 2-oxo-11-(4-(trifluoromethyl)phenyl)-11,21,51,61,7',7a1-
hexahydrospiro[indoline-3,3 1-pyrro1izine ]-21 -carboxylate 12c, 1 1 -(2,5-dimethoxypheny 1)-21 -nitro-
11,21,51,61,7',7a1-hexahydrospiro[indoline-3,31-pyrrolizin]-2-one 12m and 2-oxo-11-phenyl-1-(2-
(piperidin-1-yl)ethyl)-1 1,21 ,51,61, 7', 7a1-hexahydrospiro[indoline-3,31-pyrrolizine ]-21-carboxylic acid
13o were showing highest blood glucose lowering activity. The ED50 values for these compounds
(12c, 12m and 13o) in STZ model were found to be 62.8, 39.6 and 89.9 respectively. These
compounds were studied in detail for their anti-hyperglycemic and lipid lowering activity in db/db
mice model. Compounds 12c, 12m and 13o have shown promising anti-hyperglycemic as well as
lipid lowering activity in db/db mice model. Some of compounds of this series are selected for
preclinical studies such detail pharmacokinetic and metabolic and toxicity.
1.7. References
1. Zimmet, P.; Alberti, K. G.; Shaw, J. Nature 2001, 414, 782.
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Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
7. Diamant, M.; Heine, R. J. Drug 2003,63, 1373.
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M.; Mohr, P. Bioorg & Med Chern Lett. 2006, 16,4016--4020.
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Bioorg & Med Chern Lett. 2007, 17, 3630-3635.
19. Garrino, M.G.; Henquin, J. C. Diabetologia, 1990, 33, 145-147.
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caraway, W. T. Fundamentals of clinical chemistry, 1976, Ed Tietz NW 506-515.
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90
Chapter 2 l:_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
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Chapter 2 L::_ Design and Synthesis of Novel Spiroindoline-Heterocycle Derivatives as Anti- 2009 Diabetic A ents
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Chapter 2 Design and Synthesis of Novel Spiroindoline-Heterocycfe Derivatives as AntiDiabetic A ents
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