Table 2 Activity of ligands at H4R and selected mutants

From: Structural basis of ligand recognition and design of antihistamines targeting histamine H4 receptor

 

Histamine

Clobenpropit

Clozapine

VUF6884

JNJ7777120

hH4R

pKi

Fold (pKi)

pEC50

Fold (pEC50)

pKi

Fold (pKi)

pEC50

Fold (pEC50)

pKi

Fold (pKi)

pEC50

Fold (pEC50)

pKi

Fold (pKi)

pEC50

Fold (pEC50)

pKi

Fold (pKi)

WT

7.7 ± 0.1 (4)

 

8.4 ± 0.2 (4)

 

8.0 ± 0.1 (3)

 

8.1 ± 0.2 (6)

 

6.5 ± 0.0 (4)

 

6.6 ± 0.2 (4)

 

7.7 ± 0.1 (4)

 

7.6 ± 0.1 (4)

 

8.5 ± 0.1 (3)

 

D943.32A

ND (3)

 

ND (4)

 

ND (3)

 

ND (3)

 

ND (3)

 

ND (3)

 

ND (3)

 

ND (4)

 

ND (3)

 

D943.32N

ND (3)

 

ND (4)

 

ND (3)

 

ND (3)

 

ND (3)

 

ND (3)

 

ND (3)

 

ND (4)

 

ND (3)

 

Y953.33A

6.2 ± 0.1 (3) ****

p < 0.0001

32

↓

6.1 ± 0.1 (4) ****

p < 0.0001

200

↓

7.3 ± 0.2 (3) **

p = 0.0013

5

↓

ND (3)

 

5.7 ± 0.2 (4) ****

p < 0.0001

6

↓

ND (4)

 

6.6 ± 0.2 (4) **** p < 0.0001

12

↓

6.6 ± 0.2 (4) **

p = 0.0036

10

↓

6.7 ± 0.2 (3) ****

p < 0.0001

63

↓

E1825.46A

5.0 ± 0.1 (4) ****

p < 0.0001

501 ↓

5.4 ± 0.5 (4) ****

p < 0.0001

1000↓

6.8 ± 0.0 (3) ****

p < 0.0001

16

↓

6.1 ± 0.3 (7)

inverse agonism

100

↓

6.2 ± 0.1 (4) *

p = 0.0385

2

↓

6.4 ± 0.3 (3)

ns

p = 0.9475

2

↓

7.0 ± 0.2 (4) **

p = 0.0086

5

↓

7.6 ± 0.3 (4)

ns

p = 0.9999

1

↓

6.0 ± 0.3 (3) ****

p < 0.0001

316

↓

E1825.46Q

5.2 ± 0.1 (4) ****

p < 0.0001

316↓

5.4 ± 0.4 (3) ****

p < 0.0001

1000↓

7.2 ± 0.0 (3) ***

p = 0.0004

6

↓

8.0 ± 0.3 (3)

inverse agonism

1

↓

5.9 ± 0.0 (4) ****

p < 0.0001

4

↓

6.5 ± 0.2 (4)

ns

p = 0.9969

1

↓

6.9 ± 0.0 (4) **

p = 0.0021

6

↓

7.4 ± 0.1 (4)

ns

p = 0.9351

2

↓

7.4 ± 0.1 (3) **

p = 0.0038

13

↓

Y3196.51A

6.6 ± 0.0 (3) ****

p < 0.0001

13

↓

6.5 ± 0.2 (4) ***

p = 0.0004

80

↓

7.1 ± 0.0 (3) ***

p = 0.0001

8

↓

ND (4)

 

6.7 ± 0.0 (4)

ns

p = 0.1629

2

↑

6.4 ± 0.1 (3)

ns

p = 0.9125

2

↓

7.6 ± 0.1 (4)

ns

p = 0.9845

1

↓

7.2 ± 0.2 (4)

ns

p = 0.6146

3

↓

7.0 ± 0.0 (3) ***

p = 0.0002

32

↓

F3447.39A

4.6 ± 0.0 (3) ****

p < 0.0001

1259↓

5.3 ± 0.2 (4) ****

p < 0.0001

1259↓

5.9 ± 0.1 (3) ****

p < 0.0001

126

↓

6.0 ± 0.3 (4) ***

p = 0.0005

126

↓

5.8 ± 0.0 (4) ****

p < 0.0001

5

↓

6.1 ± 0.1 (4)

ns

p = 0.2316

3

↓

6.7 ± 0.1 (4) *** p = 0.0003

10

↓

6.7 ± 0.1 (4) *

p = 0.0121

8

↓

6.7 ± 0.2 (3) ****

p < 0.0001

63

↓

Q3477.42A

6.3 ± 0.2 (3) ****

p < 0.0001

25

↓

6.9 ± 0.1 (4) **

p = 0.0048

32

↓

7.8 ± 0.2 (3)

ns

p = 0.4132

2

↓

9.2 ± 0.2 (4)

ns

p = 0.0872

13

↑

7.7 ± 0.1 (4) ****

p < 0.0001

16

↑

8.4 ± 0.1 (4) ****

p < 0.0001

63

↑

7.3 ± 0.2 (4)

ns

p = 0.2886

3

↓

8.1 ± 0.2 (4)

ns

p = 0.1077

3

↑

6.6 ± 0.2 (3) ****

p < 0.0001

80

↓

W3487.43A

ND (3)

 

ND (4)

 

ND (3)

 

ND (4)

 

ND (3)

 

ND (3)

 

ND (3)

 

ND (4)

 

ND (3)

 
  1. Data were shown as mean ± sem of at least three independent experiments which were performed in duplicate. Binding affinity (pKi) was determined with [3H] JNJ7777120 displacement assay. Potency (pEC50) was determined with a BRET-based Gi activation biosensor. Fold decrease (↓) or increase (↑) in binding affinity or potency compared to WT H4R is indicated. Statistical difference (p < 0.05) in pKi or pEC50 for the mutants in comparison to WT H4R was analyzed for each ligand using One-way AVONA followed by Dunnett’s multiple comparison test. Statistical differences are indicated with asterisk and corresponding p-values are shown in roman. ns not significant. ND not detectable.
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