BASES TEORICAS DE LA INTERACCION FARMACO-RECEPTOR.

TRATAMIENTO Y REPRESENTACION GRAFICA DE LOS DATOS

(Segunda edicin, Noviembre de 1992)

Federico Gago Badenas

Departamento de Fisiologa y Farmacologa
Universidad de Alcal de Henares

Introduccin

2. Ensayos enzimticos.

VO=

Vmx

V mx [S]
K m + [S]

(1)

VO

Km

V
Km

V = Vmx/2:
K m + 1 = 2; = [S]
Km
[S]

(2)

1 Km 1
1
=
+
V V mx [S] V mx

Km +1
1 [S]
=
V
V mx

(3)

KI =

[E] [I]
[EI]

[EI] = [E]

(4)

[I]
KI

[Enzima total] = [ES] + [E] 1 +

[I]
KI

(1)

VI=
Km

V mx [S]
[I]
1+
+ [S]
KI

(6)

(5)

(1)

[I] = CI50, VO = 2 VI,

(6):

2 V mx [S]
CI 50
K m 1+
KI

[S]
= V mx
+ [S]
+ [S] K m

CI 50 = K I 1 +

[S]

(7)

Km

(1)

VI=
Km

V mx [S]
[I]
[I]
1+
+ [S] 1 +
K IE
K IES

(8)

[I] = CI50, VO = 2 VI

VI=

KIE =

2 V mx [S]
CI 50 + [S] 1 + CI 50
K m 1+
K IE
K IES

CI 50 =

K m + [S]
K m + [S]
K IE K IES

10

[S]
= V mx
K m + [S]

(9)

KIES

-1/Km
(9)
CI50 = KIES KIE,

CI50 = KI

(10)

KIES > KIE, Km

KIES < KIE, Km

[S] >>

Km
(9)

CI 50 =

CI 50 =

Km/[S]

<<

KIE/KIES

Km/[S]

CI50 = KIES

K IE
K m + K IE
[S] K IES

(11)

(13)

(7)

11

1
Km 1 + 1
[S] K IE K IES

(12)

(11)

Vmx

1 + [I]/KI

Km

1/Vmx
-1/Km

CI50 = KI

[S] >> Km

1 + [I]/KI

12

Figura 7

Figura 6

KI
[I]

1/V

KI

(1)
(3)

V = V mx - K m

V
[S]

V V/[S]

(3)

[S]

[S]/V

13

[S]

V Vmx

(14)

3. Respuestas graduales y teora ocupacional.

log [F1] - log [F2]

14

(15)

k1 [F] [R] = k2 [FR]

k1

k2

[FR]

[R]

K 2 = [F] [R] =
K D = Constante de disociacin en el equilibrio
[FR]
K1

(16)

[Rt]
[Rt] = [FR] + [R]

KD =

[R] = [Rt] - [FR]

[F] ([ Rt ] - [FR])
[ ]
= [F] Rt - 1
[FR]
[FR]

(17)

KD
[FR] = [Rt]/2, KD = [F]

Ka = K1/K2

FR
F
F
=r=
= Ka
Rt
1+ Ka F
KD + F

(18)

KA

KD

15

[F]

[F]por KD

E
F

E mx

[F]
[FR]
=
K D + [F] [ Rt ]

(19)

Emx
E(KD + [F]) = Emx [F]

E
Emx E

[F]
KD

(20)

[FR] ([KD]+ [F]) = [Rt] [F]

[FR]
[F]
=
[ Rt ] - [FR] K D

16

(21)

log([FR]/([Rt]-[FR]))

log(E/Emx-E)

log[F]

log[F]

Emx = K3 [Rt]

[F] = [F]total
(19):
[F]
E = E mx
K D + [F]

(22)

(1)

E = Emx/2
KD/[F] + 1 = 2; KD = [F]

(23)

(21)
KD
+1
K
1 [F]
1
1
=
= D
+
E
E mx
E mx [F] E mx

(3)

17

(24)

(19)

(19)

E
E mx

[FR]
[ Rt ]
18

(25)

(22):
E
E mx

0 <

[F]
K D + [F]

(26)

< 1

(24)
E m x K D 1
1
=
+
E
[F]

1/[F]

(27)

Emx/E = 1/

Emx/E

1/[F] = -1/KD

19

S=

e [FR]
e [F]
=
[ Rt ]
[F] + K D

20

(28)

E
E mx

= f (S)= f

e [F]
[F] + K D

21

(29)

4. Respuestas equivalentes.

Respuesta =

[F ]1 K a
[F ] 2 K a
=
1 + [F ]1 K a [F ] 2 K a + [A] K ant + 1

d=

[F ] 2
= [A] K ant + 1
[F ]1

(28)

Kant

Kant
(28)

Kant
Kant

22

K 1 [F] ([ Rt ] - [AR] - [FR])= K 2 [FR]

Ka =

[FR]
K1 =
K 2 [F] ([ Rt ] - [AR] - [FR])

K a [F] =

[FR]
([ Rt ] - [AR] - [FR])

(29)

[AR]
([ Rt ] - [AR] - [FR])

(30)

K ant [A] =

[AR]= K ant [A] ([ Rt ] - [AR] - [FR])

[AR]= K ant [A] ([ Rt ] - [FR])- K ant [A] [AR]

[AR] (1+ K ant [A]) = K ant [A] ([ Rt ] - [FR])

[AR] =

(29)

([ Rt ] - [FR]) K ant [A]

1 + K ant [A]

(30)
[F] K a [FR]
[FR] (1 + [A] K ant )
=
=
[A] K ant [AR] ([ Rt ] - [FR]) [A] K ant

23

[F] K a =

[FR] (1 + [A] K ant )

[FR]
=
(1 + [A] K ant )
([ Rt ] - [FR])
[ Rt ] - [FR]

[F]2
[A]
[F]1

(17)

[F ]1 K a =

[FR]
[ Rt ] - [FR]

[FR]
[F]1

[A]

[F]2
[F ] 2
= d = [A] K ant + 1
[F ]1

K ant [A] = 1

log K ant = - log [A] = pA2

24

(28)

log K ant + log [A] = 0

(31)

d - 1 = [A ] x K ant
log(d - 1) = log [A ] x + log K ant

(32)

log(d - 1) = - pAx + pA2 = pA2 - pAx

d = 10, pA2 - pA10 = log 9 = 0.95

log (d - 1) = log [B] - log K B

log(d-1)

(33)

-log[A]

25

5. Ensayos de unin de ligandos.

26

[LR]
log[L]

[LR]/[L]

[LR]

27

R
LR
LR
= t
, de donde:
L
KD

(21)

[LR] [ Rt ] 1
=
[LR]
[L]
Kd Kd

[LR]/[L]

(34)

[LR]

[Rt]= OC
[LR]= OB

OC OB .

(35)

[LR]
k [L]
=
[ Rt ] 1 + k [L]

(35)

[LR] n1 k 1 [L]
[L]
=
+ n2 k 2
[ Rt ] 1 + k 1 [L] 1 + k 2 [L]

28

(36)

n1 n2
(36)

(36)

kd1 kd2

[ ]
[ ]
[LR]
= Rt 1 + Rt 2
[L]
[L] + k d 1 [L] + k d 2

OP = OP 1 + OP 2 .

29

(37)

"n x m"

30

6. Experimentos de desplazamiento de ligandos.

KI

(7)

CI 50 = K I 1 +

[L]
KD

31

(37)

KI =

CI 50
[L]
1+
KD

32

(38)

33

7. Receptores de reserva.

34

35

36

8. Respuestas cuantizadas: efectos de todo-o-nada.

37

s=
39

d
n -1

(39)

x ( (x - x ) = 0 ;

38

(x - x )2 es positivo)

N2

P=
N1

1
2

-1/2 N 2

dN

39

N = N = 0

x= x

Ns

40

(40)

ES DE50 =

2s
2n

LC DE50 = DE 50 t s

41

(41)

(42)

42

9. Margen de seguridad. Evaluacin del ndice teraputico.

REFERENCIAS SELECCIONADAS.

0
1
2
3
4

2,6737
2,9463
3,1192
3,2493

20
21
22
23
24

4,1584
4,1936
4,2278
4,2612
4,2937

40
41
42
43
44

4,7467
4,7725
4,7981
4,8236
4,8490

60
61
62
63
64

5,2533
5,2793
5,3055
5,3319
5,3585

80
81
82
83
84

5,8416
5,8779
5,9154
5,9542
5,9945

5
6
7
8
9

3,3551
3,4452
3,5242
3,5949
3,6592

25
26
27
28
29

4,3255
4,3567
4,3872
4,4172
4,4466

45
46
47
48
49

4,8743
4,8996
4,9247
4,9498
4,9749

65
66
67
68
69

5,3853
5,4125
5,4399
5,4677
5,4959

85
86
87
88
89

6,0364
6,0803
6,1264
6,1750
6,2265

10
11
12
13
14

3,7184
3,7735
3,8250
3,8736
3,9197

30
31
32
33
34

4,4756
4,5041
4,5323
4,5601
4,5875

50
51
52
53
54

5,0000
5,0251
5,0502
5,0753
5,1004

70
71
72
73
74

5,5244
5,5534
5,5828
5,6128
5,6433

90
91
92
93
94

6,2816
6,3408
6,4051
6,4758
6,5548

15
16
17
18
19

3,9636
4,0055
4,0458
4,0846
4,1221

35
36
37
38
39

4,6147
4,6415
4,6681
4,6945
4,7207

55
56
57
58
59

5,1257
5,1510
5,1764
5,2019
5,2275

75
76
77
78
79

5,6745
5,7063
5,7388
5,7722
5,8064

95
96
97
98
99

6,6449
6,7507
6,8808
7,0537
7,3263

Tabla 2. Distribucin t de Student.

grados
de
libertad

90%

95%

(P=0,1)

(P=0,5)

(P=0,01)

1
2
3
4
5

6,314
2,920
2,353
2,132
2,015

12,706
4,303
3,182
2,776
2,571

63,657
9,925
5,841
4,604
4,032

6
7
8
9
10

1,943
1,895
1,860
1,833
1,812

2,447
2,365
2,306
2,262
2,228

3,707
3,499
3,355
3,250
3,169

11
12
13
14
15

1,796
1,782
1,771
1,761
1,753

2,201
2,179
2,160
2,145
2,131

3,106
3,055
3,012
2,977
2,947

16
17
18
19
20

1,746
1,740
1,734
1,729
1,725

2,120
2,110
2,101
2,093
2,086

2,921
2,898
2,878
2,861
2,845

21
22
23
24
25

1,721
1,717
1,714
1,711
1,708

2,080
2,074
2,069
2,064
2,060

2,831
2,819
2,807
2,797
2,787

26
27
28
29
30

1,706
1,703
1,701
1,699
1,645

2,056
2,052
2,048
2,045
1,960

2,779
2,771
2,763
2,756
2,576

99%

PROBLEMA 1.

10

PROBLEMA 2.

PROBLEMA 3.

PROBLEMA 4.

PROBLEMA 5.

10

PROBLEMA 6.

11

PROBLEMA 7.

12

INDICE DE MATERIAS.

Pg.

1.
2.
3.
4.
5.

Ecuacin de Michaelis-Menten ...................

Ecuacin y representacin de Lineweaver-Burk....
Inhibicin competitiva..........................
Inhibicin no-competitiva .....................
Ecuaciones de Cheng-Prusoff:
5.1. Inhibidores competitivos...................
5.2. Inhibidores no-competitivos................
6. Representacin de Dixon.........................
7.
8.
9.
10.
11.
12.
13.
14.
15.

Representaciones de Eadie-Hoffstee y Hanes-Woolf

Potencia relativa de los frmacos...............
Aplicacin de la ley de accin de masas.........
Ecuacin de Clark...............................
Ecuacin y representacin de Hill...............
Eficacia y actividad intrnseca.................
Agonistas parciales o "dualistas"...............
Respuestas equivalentes (Gaddum)................
Representacin de Schild. Concepto de pA2.......

16. Ecuacin y representacin de Scatchard..........

17. Tcnica de Rosenthal para un sistema con ms de
un receptor......................................
18. Determinacin de la CI50 y la Ki en un ensayo de
fijacin ........................................
19. Capacidad de reserva de los receptores...........
20. Sistemas de amplificacin........................
21. Respuestas cuantizadas todo-o-nada...............
22. Probits..........................................

6
7,18
8
10
9
10-11
13
13
13
13-14
15
17
18
19
22
24-25
26-27
29-30
31
34
34
37
40

23. Clculo de la DE50: mtodo de Miller y Tainter... 41

24. Margen de seguridad. Evaluacin del ndice
teraputico...................................... 43
29. Referencias seleccionadas........................ 44
26. Tabla de probits................................. 45
27. Distribucin t de Student........................ 46
28. Ejercicios numricos ............................ 47-51

13