1.      摘要：目的：初步测定解毒维康片对人体白血病K562及小鼠Lewis肺癌（皮下接种）模型的实验抗肿瘤疗效。方法：取K562实体瘤的第二代肿瘤，或Lewis肺癌分别以生理盐水稀释成约5至6×10⁷/ml癌细胞匀浆，0.2ml/鼠，皮下接种，次日按方案给药，实验结束取肿瘤与阴性对照组相比计算各组的抑瘤率。人体白血病K562模型还需动态检测各组荷瘤鼠肿瘤及体重的生长变化。结果：解毒维康片对人体白血病K562以5g/kg/d、2.5g/kg/d、1.25g/kg/d剂量ig×14bid，两次抑瘤率分别为48.00%、36.00%、26.64%及47.12%、37.35%、29.32%。对小鼠Lewis肺癌，以5g/kg/d、2.5g/kg/d、1.25g/kg/d剂量ig×10qd及5g/kg/d剂量ig×10bid两次抑瘤率分别为45.87%、38.53%、30.73%、49.08%及47.15%、39.84%、31.30%、51.63%。结论：解毒维康片以其最大耐受量的1/4、1/8、1/16设定为高、中、低三个剂量灌胃给药对人体白血病K562（皮下接种）裸鼠实体瘤模型及对小鼠Lewis肺癌（皮下接种）模型均具有中等度的抑瘤效果；但相同剂量每日2次分隔给药比每日1次给药效果更为明显。

    ABSTRACT  purpose:  To determine the antineoplastic effectiveness of Jiedu Weikang Tablets on human leukemia K562 and Lewis pulmonary carcinoma ( subcutaneous injection ) in mice. Methods:  To remove the second generation of solid tumor K562 or Lewis pulmonary carcinoma and then the tumor mass was diluted by physiological saline into tumor cells homogenate ( 5 x 10’7 to 6 x 10’7/ ml ) and the homogenate at dose of 0.2 ml/mouse was injected into mice subcutaneously. At the next day mice orally administered the drug according to the scheme for administration of test drug. The tumor mass was removed and measured in test drug and control group and compared with that in control group at the end of test, and inhibitory rate for tumor in test drug and control groups was calculated. The tumor weight and body weight were dynamically measured in mice loaded human leukemia K562. Results:  Jiedu Weikang Tablets at dose of 5.0 g/kg/d, 2.5 g/kg/d and 1.25 g/kg/d ( twice a day ) were orally administered by mice loaded human leukemia K562 for 14 days and the inhibitory rate for tumor was 48.00%, 36.00 %, 26.64 % ( the first test ) and 47.12 %, 37.35 %, 29.32 % ( the second test ) respectively. Jiedu Weikang Tablets at dose of 5.0 g/kg/d, 2.5 g/kg/d and 1.25 g/kg/d or at dose of 5.0 g/kg/d ( twice a day ) were orally administered by mice loaded Lewis pulmonary carcinoma for 10 days and the inhibitory rate for tumor was 45.87 %, 38.53 %, 30.73 %, 49.08 % ( the first test ) and 47.15 %, 39.84 %, 31.30 %, 51.63 % ( second test ) respectively.  Conclution:  The high, medium and low dose of Jiedu Weikang Tablets corresponded to 1/4, 1/8 and 1/16 of maximum oral dose respectively and then Jiedu Weikang Tablets at high, meddium and low dose were oral administered by nude mice  loaded human leukemia K562 or mice loaded Lewis pulmonary carcinoma, and Jiedu Weikang Tablets had moderate antineoplastic effectiveness. The therapeutic effectiveness of Jiedu Weikang Tablets at same dose ( divided into two administrations ) was better than that at same dose ( single administration ).

2.      目的：初步测定解毒维康片对人体白血病K562及小鼠Lewis肺癌（皮下接种）模型的实验抗肿瘤疗效。

    Purpose:  To determine the antineoplastic effectiveness of Jiedu Weikang Tablets in mice loaded human leukemia K562 and mice loaded Lewis pulmonary carcinoma ( subcutaneous injection ).  

3.      受试药物。　　Test drug:  

3.1 名称：解毒维康片片剂。　Name:  Jiedu Weikang Tablets

3.2 提供单位：西安恒生堂制药有限公司。

Providing unit:  Xi’an Hengshengtang Pharmaceutical Co., Ltd

3.3 药品批号：030101。　Batch:  030101

3.4 配制方法：取解毒维康片片剂一定量，置于研钵中，捣碎研细，加入少量吐温80湿润，以0.5%CMC-Na溶液研磨，使成混悬液，再以0.5%CMC-Na溶液稀释配制成所需的各档试液。每鼠每次灌胃体积为0.5ml。

Preparation of test drug:  Jiedu Weikang Tablets were grinded into powder in mortar and the powder moisted by Tween 80 was grinded by 0.5 % CMC-Na into suspension. The suspension was diluted into liquid of various concentration by 0.5 % CMC-Na. Each oral administered volume was 0.5 ml/mouse.

4.      实验材料。　Experimental materials:  

4.1 溶剂：0.5%CMC-Na溶液　Solvent:  0.5 % CMC-Na liquid

4.2 阳性对照品：注射用环磷酰胺，上海华联制药厂出品。

Positive drug: Cyclophosphamine for injection, Shanghai Lianhua Pharmaceutical Factory.

4.3 瘤源：人体白血病K562实体瘤的第二代肿瘤及小鼠Lewis肺癌，由上海医药工业研究院药理室体内传代维持。

Strain:  The second generation of solid human leukemia K562 ( solid tumor ) and Lewis pulmonary carcinoma were generated in Department of Pharmacology, Shanghai Academy of Medicinal Industry.

5.      实验动物。　Experimantal animals:

5.1 来源：裸小鼠及C57BL/6小鼠均由上海中科院实验动物中心提供，合格证号：中科动管第005号。

Origin and strain:  Nude mice and C57BL/6 mice were supplied by Experimental Animal Center, Chinese Academy of Sciences and certificate of quality is zhongkedongguan No. 005.

5.2 体重：17至21克。Body weight:   17 to 21 g

5.3 性别：雌性。Sex:  Female

5.4 动物数：试验组及阳性对照组裸小鼠每组6只小鼠；C57BL/6小鼠每组10只小鼠，阴性对照各为两组。Number of animals:  Six nude mice in test drug and positive drug groups, and 10 C57BL/6 mice in test drug and positive drug groups and there were two groups in control group.

6.      剂量设置：根据小鼠灌胃最大耐受量的1/4、1/8、1/16设计为高、中、低三个剂量，即5g/kg/d、2.5g/kg/d、1.25g/kg/d剂量。

Dosage:  The high, medium and low dose were 5.0 g/kg/d, 2.5 g/kg/d and 1.25 g/kg/d ( corresponded to 1/4, 1/8 and 1/16 of maximum oral dose in mice ) respectively.

7. 给药方案：灌胃给药，每天1次或分隔为2次，连续10天或14天。

Scheme for administration:  Oral administration, once or twice a day for 10 or 14 days.

8. 试验对照：阴性对照给以与试验组等体积的0.5%CMC-Na溶液，给药方案为灌胃给药，每天1次，连续14天。阳性对照环磷酰胺（CTX）30mg/kg，腹腔或静脉给药，每天1次，连续7天。

Control test:  Same volume of 0.5 % CMC-Na liquid ( once a day ) was orally administered for 14 days in control group and positive drug ( CTX ) at dose of 30 mg/kg ( once a day ) was given for 7 day peritoneally or intravenously.

9.      试验主要步骤：　Main steps:

无菌条件下取K562人体白血病实体瘤的第二代肿瘤，以生理盐水1:5进行稀释成约5至6×107/ml癌细胞匀浆，0.2ml/鼠，分别皮下接种，接种24小时后按实验设计方案给药，每周动态检测各组荷瘤鼠肿瘤（采用卡尺测量每个肿瘤的长、短径。肿瘤体积=短径×长径/2）及体重的生长变化。结束时取出肿瘤称重与阴性对照组相比，按下式计算各组的抑瘤率：

To take the second generation of human leukemia K562 ( solid tumor ) under sterile condition and the solid tumor was diluted by physiological saline ( tumor cell : saline =1 : 5 ) into tumor cells homogenete ( 5 x 10’7 to 6 x 10’7/ml ), and then the tumor cells homogenete at dose of 0.2 ml/mouse was injected subcutaneously. The test drug was administered 24 hours after transplantation according to the scheme for administration and the tumor mass and body weight were dynamically measured in tumor-loaded mice of various groups weekly. And The tumor mass ( long and short diameter ) was measured by callipers ( tumor volume = short diameter’2 x long diameter/2 ) and the tumor mass was removed and its weight was measured at the end of test, and their tumor mass weight in drug group was compared with that in control group. Then the inhibitory rate for tumor was calculated as follow formula.

抑瘤率%=[（给药组肿瘤体积－对照组肿瘤体积）/对照组肿瘤体积]×100

inhibitory rate for tumor = tumor volume in control group - tumor volume in drug group  x 100 tumor volume in control group

实验用裸小鼠所用的饲料、垫料、笼具及接触的器械等均应高压消毒后使用，裸小鼠置天层流架中SPF条件饲养。

Food, bedding material , cages, contacted apparatus and others for nude mice were sterilized by high pressure and nude mice were on laminar flow framework and were raised according to SPF condition.

小鼠Lewis肺癌操作相同，但不作肿瘤及体重的动态观察。

Experiment for mice loaded Lewis pulmonary carcinoma was same as above-mentioned steps, but the tumor mass and body weight did not be observed dynamically.

1.      试验结果：解毒维康片对人体白血病K562以5g/kg/d、2.5g/kg/d、1.25g/kg/d剂量ig×14bid，两次抑瘤率分别为48.00%、36.00%、26.64%及47.12%、37.35%、29.32%，详见表1、2图1及2。动态检测各组荷瘤鼠肿瘤及体重的生长变化详见表3、4、5、6及图3、4、5、6。解毒维康片采用以上相同剂量5g/kg/d、2.5g/kg/d、1.25g/kg/d剂量ig×10qd及5g/kg/d剂量ig×10bid对小鼠Lewis肺癌，两次抑瘤率分别为45.87%、38.53%、30.73%、49.08%及47.15%、39.84%、31.30%、51.63%，详见表7、8，图7。

 Rusults:

     Jiedu Weikang Tablets at dose of 5.0 g/kg/d, 2.5 g/kg/d 1.25 g/kg/d ( twice a day ) were orally administered by mice loaded human leukamia K562 for 14 days and the inhibitory rate for tumor was 48.00 %, 36.00 %, 26.64 % ( the first test ) and 47.12 %, 37.35 %, 29.32 % ( the second test ) respectively. See Table 1, 2 and Figure 1, 2. The changes of tumor mass and body weight in tumor-loaded mice after dynamic measurement were in Table 3, 4, 5, 6 and Figure 3, 4, 5, 6. Jiedu Weikang Tablets at dose of 5.0 g/kg/d, 2.5 g/kg/d, 1.25 g/kg/d ( once a day ) and 5.0 g/kg/d ( twice a day ) were orally administered by mice loaded Lewis pulmonary carcinoma for 10 days and the inhibitory rate for tumor was 45.87 %, 38.53 %, 30.73 %, 49.08 % ( the first test ) and 47.15 %, 39.84 %, 31.30 %, 51.63 % ( the second test ) respectively. See Table 7, 8 and Figure 7.

2.      试验小结和讨论：Conclusion and discussion

11.1 解毒维康片以其最大耐受量的1/4、1/8、1/16为高、中、低三个剂量灌胃给药对人体白血病K562（皮下接种）异种接种于裸小鼠实体瘤模型及对小鼠Lewis肺癌（皮下接种）模型均具有中等度的抑瘤效果。但相同剂量每日2次分隔给药比每日1次给药效果更为明显。

Jiedu Weikang Tablets at high, medium and low dose ( corresponding to 1/4, 1/8 and 1/16 of maximum oral dose ) were orally administered by nude mice loaded human leukemia K562 and mice loaded Lewis pulmonary carcinoma subcutaneously and had moderate inhibitory effect on tumor, and the therapeutic effectiveness of same dose ( divided into two times ) was better than that of the same dose ( once a day ).

11.2 解毒维康片为中药复方制剂，动物肿瘤模型治疗剂量的设计是根据小鼠的最大耐受量及临床每天推荐剂量推算，但是由于样品毒性小，用药量相对为大，而小鼠胃容量又有限，考虑每天1次灌胃量过多会影响胃的正常蠕动，非但影响小鼠对食物的正常摄入，也妨碍药物的吸收，故将每天1次灌胃量分隔成2次给药，实际试验的结果证明，在每天总剂量相同的情况下灌胃分隔成2次给药方案的效果要优于1次给药的方案，解毒维康片对Lewis肺癌的抑瘤率要超过5个百分点。实验过程还发现给药后动物的体重增长与对照相似，提示本品毒性较低。

     Jiedu Weikang Tablets is a compound recipe of traditional Chinese drugs and the therapeutic dose for animals was designed according to maximum oral dose in mice and recommended clinical daily dose. The test drug was low toxic effect and larger administered volume, and mouse had limited gastric capacity and excessive volume of test drug ( once a day ) maybe influence normal peristalsis and normal intake of food and absorption of test drug in mice. So the daily dose was divided into two parts and the divided dose was orally administered twice a day. Experimental results proved that the therapeutic effectiveness of same daily dose ( divided to two times ) was better than that of same daily dose ( once a day ), and Jiedu Weikang Tablets for two administrations scheme enhanced five percentage points of the inhibitory rate for tumor in mice loaded Lewis pulmonary carcinoma.  The increase of body weight in test drug group was similar with that in control group and it is considered that toxic action of Jiedu Weikang Tablets is lower.

表1.解毒维康片对K562人体白血病（腋皮下接种）异种接种于裸小鼠的疗效之一。

Table 1.  Effect of drug on tumor mass weight in nude mice loaded K562 ( first test )

试验组与阴性对照组相比，***表示P值＜0.01。Note: *** P < 0.01, compared with control group

表2.解毒维康片对K562人体白血病（腋皮下接种）异种接种于裸小鼠的疗效之二。

Table 2.  Effect of drug on tumor mass weight in nude mice loaded K562 ( second test )

试验组与阴性对照组相比，***表示P值＜0.01。Note:  *** P < 0.01, compared with control group 

解毒维康片样品对K562实体的疗效试验

Therapeutic effectiveness of drug in nude mice loaded leukemia K562

阴性对照 Control     相应溶剂ig×14bid

阳性对照CTX    30mg /kg ip×7pd

解毒维康片 Jiedu Weikang   5g/kg/天ig×14bid

解毒维康片  Jiedu Weikang    2.5 g/kg/天ig×14bid

解毒维康片 Jiedu Weikang       1.25 g/kg/天ig×14bid                 

解毒维康片样品对K562实体的疗效试验

Therapeutic effectiveness of drug on tumor mass weight in nude mice loaded K562

阴性对照 Control       相应溶剂ig×14bid

解毒维康片Jiedu Weikang      5g/kg/天ig×14bid

解毒维康片 Jiedu Weikang     2.5 g/kg/天ig×14bid

解毒维康片Jiedu Weikang      1.25 g/kg/天ig×14bid

阳性对照CTX    30mg /kg ip×7pd

表3.解毒维康片对K562人体白血病（腋皮下接种）疗效试验中肿瘤动态变化之一

Table 3.  Effect of drug on tumor volume in mice loaded K562  (x ± s) (first test)

Tumor volume ( cm’3 )     day after transplantation

解毒维康片对K562人体白血病实体瘤疗效试验中肿瘤动态变化之一

Dynamic change on effect of drug on tumor volume in mice loaded K562

阴性对照Control           0.5%CMC-Na     ig×14bid

阳性对照CTX      30mg /kg/d        ip×7pd

CH-001 Jiedu Weikang           5g /kg/d          ig×14bid

CH-001 Jiedu Weikang           2.5g /kg/d         ig×14bid

CH-001 Jiedu Weikang           1.25g /kg/d        ig×14bid

表4.解毒维康片对K562人体白血病（腋下皮接种）疗效试验中荷瘤鼠体重动态变化之一

Table 4.  Effect of drug on body weight inmice loaded K562 (x ± s) ( first test )

Body weight of mice loaded K562 (g)  day after transplantation  

解毒维康片对K562人体白血病（腋皮下接种）疗效试验中荷瘤鼠体重动态变化之一

Dynamic change on effect of drug on body weight in mice loaded K562

阴性对照Control           0.5%CMC-Na     ig×14bid

阳性对照CTX      30mg /kg/d        ip×7pd

CH-001 Jiedu Weikang         5g /kg/d          ig×14bid

CH-001 Jiedu Weikang           2.5g /kg/d         ig×14bid

CH-001 Jiedu Weikang           1.25g /kg/d        ig×14bid

表5.解毒维康片对K562人体白血病（腋皮下接种）疗效试验中肿瘤动态变化之二

Table 5.  Effect of drug on tumor volume in mice loaded K562 (x ± s) (second test)

Tumor volume (cm’3)    day after transplantation               

解毒维康片对K562人体白血病实体瘤疗效试验中肿瘤动态变化之二

Dynamic change on effect of drug on tumor volume in mice loaded K562

阴性对照 Control          0.5%CMC-Na     ig×14bid

阳性对照CTX      30mg /kg/d        ip×7pd

CH-001 Jiedu Weikang           5g /kg/d          ig×14bid

CH-001 Jiedu Weikang           2.5g /kg/d         ig×14bid

CH-001 Jiedu Weikang          1.25g /kg/d        ig×14bid

表6.解毒维康片对K562人体白血病（腋皮下接种）疗效试验中荷瘤鼠体重动态变化之二

Table 6.  Effect of drug on body weight in mice loaded K562 (x ± s) ( first test)

ody weight of mice loaded K562 (g)      day after transplantation

解毒维康片对K562人体白血病（腋皮下接种）疗效试验中荷瘤鼠体重动态变化之二

Dynamic change on effect of drug on body weight in mice loaded K562

阴性对照 Control          0.5%CMC-Na     ig×14bid

阳性对照CTX      30mg /kg/d        ip×7pd

CH-001  Jiedu Weikang          5g /kg/d          ig×14bid

CH-001 Jiedu Weikang           2.5g /kg/d         ig×14bid

CH-001 Jiedu Weikang           1.25g /kg/d        ig×14bid

表7.解毒维康片对小鼠Lewis肺癌（腋皮下接种）模型的疗效之一

Table 7.  Effect of drug on tumor weight in mice loaded Lewis pulmonary carcinoma (first test)

试验组与阴性对照组相比，***表示P值＜0.01。Note:  *** P < 0.01, compared with control group

表8. 解毒维康片对小鼠Lewis肺癌（腋皮下接种）模型的疗效之二

Table 8.  Effect of drug on tumor weight in mice loaded Lewis pulmonary carcinoma (second test)

试验组与阴性对照组相比，***表示P值＜0.01。Note:  *** P < 0.01, compared with control group

Therapeutic effectiveness of drug on tumor mass in mice loaded Lewis pulmonary carcinoma

解毒维康片样品对Lewis肺癌实体的疗效试验

Figure 7.  Effect of drug on tumor mass in mice load

阴性对照相Control         相应溶剂ig×10qd

CH-001  Jiedu Weikang        5g /kgig×10qd

CH-001 Jiedu Weikang        2.5g /kgig×10bid

CH-001 Jiedu Weikang        2.5g /kgig×10qd

CH-001 Jiedu Weikang         1.25g /kgig×10qd

阳性对照相CTX  30mg /kg ip×7pd

来源：恒生堂