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To disable access to a custom application for a group of users, using profiles is the most appropriate method. Profiles in Salesforce define a user's permissions and access levels within the platform. Each user is assigned a profile that determines their capabilities, including access to specific applications.

When a system administrator wants to restrict access to a custom application, they can modify the settings in the profile associated with the group of users. By unchecking the permission to access the custom application in the corresponding profile, the administrator effectively disables access for that group. This method allows for clear and structured management of user access based on their roles and responsibilities within the organization.

Other options like Web Tabs, Page Layouts, and Sharing Rules do not effectively manage application-level access for groups of users. Web Tabs are used to create custom tabs for navigation within an application, Page Layouts dictate how records are displayed but do not control application access, and Sharing Rules are designed for record-level sharing, not application access. Hence, profiles serve as the foundational means of managing user access to applications in Salesforce.